Derivatives of physiologically active substance K-252

ABSTRACT

The present invention relates to novel derivatives of K-252, (8R*, 9S*, 11S*)-(-)-9-hydroxy-9-methoxycarbonyl-8-methyl-2,3,9,10-tetrahydro-8,11-epoxy-1H,8H, 11H-2,7b,11a-triazadibenzo[a,g]cycloocta-[c,d,e,]trinden-1-one, and the compounds are represented by formula (I): ##STR1## wherein W 1 , W 2 , R 1 , R 2 , R 3 , R 4 , X and Y represent various substituents. The compounds are physiologically active substances that inhibit protein kinase C and exhibit an antitumor activity, and are useful as medicines.

TECHNICAL FIELD

The present invention relates to novel compounds which inhibit proteinkinase C (hereinafter referred to as C-kinase) and have pharmacologicalactivities such as anti-tumor activity.

BACKGROUND ART

C-kinase is a protein kinase which is activated depending uponphospholipids and calcium and widely distributed over tissues and organsin the living body. It has been reported that this enzyme plays anextremely important role in cell membrane receptor transductionmechanism in which many hormone, neurotransmitters, etc. are concerned.As examples of physiological response induced by signal transductionsystem in which C-kinase participates, there have been reportedserotonin release from platelets, lysosomal enzyme release andaggregation. superoxide formation and lysosomal enzyme release fromneutrophil leukocytes, epinephrine release from adrenal medulla,secretion of aldosterone from renal glomerulus, secretion of insulinfrom Langerhans' islet, histamine release from mast cells, acetylcholinerelease from ileum, contraction of vascular smooth muscle, and the like.C-kinase is also supposed to be concerned in cell growth andcarcinogenetic mechanism [Y. Nishizuka, Science, 225, 1365 (1984);Rasmusen et al., Advance in Cyclic Nucleotide and ProteinPhosphorylation Research, vol. 18, p. 159, edited by P. Greengard and G.A. Robinson, Raven Press, New York, 1984]. It is expected that a widevariety of diseases such as diseases of the circular system,inflammatory diseases, allergy an tumor can be prevented or treated byartificially inhibiting C-kinase activity by the use of inhibitors, etc.

On the other hand, it has been found that antipsychotic drugs such astrifluoperazine and chlorpromazine, dibenamine and tetracaine which areknown as local anesthetics, calmodulin inhibitor W-7[N-(6-aminohexy)-5-chloro-1-naphthalenesulfonamide], etc. posssessC-kinase inhibitory activity, but the C-kinase inhibitory activity islow in any of these drugs [Y. Nishizuka et al., J. Biol. Chem., 255,8378 (1980); R. C. Schatzman et al., Biochem. Biophys. Res. Commun., 98,669 (1981); B. C. Wise et al., J. Biol. Chem., 257, 8489 (1982)].

K-252 and KT-5556 represented by the following formula are known (withK-252, see Japanese Published Unexamined Patent Application No. 41489/85and U.S. Pat. No. 4,555,402; with KT-5556, see Japanese PublishedUnexamined Patent Application No. 176531/86). ##STR2## K-252: R_(A)=CH₃, R_(B) =H KT-5556: R_(A) =H, R_(B) =H

In Japanese Published Unexamined Patent Application No. 41489/85, it isdescribed that K-252 has activity to inhibit histamine release andanti-allergic activity. In Japanese Published Unexamined PatentApplication No. 176531/86, it is described that KT-5556 has activity toinhibit histamine release. Further, compounds that are assumed to beidentical with K-252 or KT-5556 have been reported as antibacterialsubstances [M. Senzaki et al., J. Antibiotics, 38 (10), 1437 (1985)]. Inthis publication, a compound of the above formula wherein R_(A) =CH₃ andR_(B) =Ac is also disclosed.

Furthermore, Staurosporine having the following structure andantibacterial activity is known as a compound having a structurerelatively akin to that of K-252 [S. Omura et al., J. Antibiotics, 30(4), 275 (1977), A. Furusaki et al., J. Chem. Soc. Chem. Commun., 800(1978), Japanese Published Unexamined Patent Application No. 185719/85].##STR3##

DISCLOSURE OF THE INVENTION

According to the present invention, there are provided novel derivativesof K-252 represented by formula (I) and pharmacologically acceptablesalts thereof. ##STR4##

In the formula, R¹ represents hydrogen, methyl, hydroxy, hydroxymethyl,lower alkoxy, bromine, chlorine or --NR⁵ R⁶ (wherein either R⁵ or R⁶ ishydrogen and the other is hydrogen, carbamoyl or loweralkylaminocarbonyl, or both are lower alkyl) and R³ is hydrogen, or R¹and R³ are the same and represent hydrosy, lower alkoxy or amino. R² ishydrogen or amino, and R⁴ is hydrogen, chlorine, carbamoyl, lower alkyl,amino or --CH₂ CH₂ R⁷ (wherein R⁷ is bromine, amino, di-loweralkylamino, hydroxy or hydroxysubstituted lower alkylamino).

W₁ and W₂ are hydrogen or both are combined together to representoxygen. X is hydrogen, formyl, lower alkoxycarbonyl, ##STR5## (whereinR⁸ and R⁹ are independently hydrogen, lower alkyl or hydroxy-substitutedlower alkyl; or R⁸ is hydrogen and R⁹ is hydroxy), CH₂ A {wherein A ishydroxy, azido, lower alkylthio, lower alkylsulfenyl, ##STR6## [whereineither R¹⁰ or R¹¹ is hydrogen and the other is hydrogen, lower alkyl,allyl, carboxylic acid-substituted lower alkyl, dihydroxysubstitutedlower alkyl, a residue of an α-amino acid in which the hydroxy of thecarboxylic acid is removed or lower alkoxycarbonyl-substituted loweralkyl; or both are lower alkyl or chlorine-substituted lower alkyl; orR¹⁰ and R¹¹ are combined together to form --CH₂ CH₂ BCH₂ CH₂ -- (whereinB is --CH₂ --, --NH--, --S-- or --O--)], --N=CH--NMe₂ (wherein Me ismethyl), --OCOCH₂ CH₂ CO₂ H or ##STR7## or --CH═NR¹² (wherein R¹² ishydroxy, amino, guanidino or 2-imidazoylamino).

Y is hydroxy or carbamoyloxy; or X and Y are combined together to form,as --X--Y--, O=, --CH₂ O--, --CH₂ OCOO--, --CH₂ --O--CS--O--, --CH₂--NR¹³ --CO--O-- (wherein R¹³ is hydrogen, lower alkyl, allyl,formylmethyl, --CH₂ CH(OH)--CH₂ OH, ##STR8## --CH₂ --NH--CS--O--, --CH₂--O--SO--O-- or ##STR9## (wherein R¹⁴ is lower alkyl or loweralkylthio).

With the proviso that W₁ and W₂ are combined together to representoxygen, R¹, R² and R³ are all hydrogen. When R⁴ is lower alkyl, amino or--CH₂ CH₂ R⁷, W₁ and W₂ are combined together to represent oxygen. WhenY is carbamoyloxy, R¹, R², R³, W₁ and W₂ are all hydrogen, R⁴ iscarbamoyl and X is lower alkoxycarbonyl. When R⁴ is chlorine, R¹, R²,R³, W₁ and W₂ are all hydrogen and X is lower alkoxycarbonyl. When X ishydrogen, formyl, ##STR10## (wherein R^(8a) and R^(9a) are independentlyhydrogen, lower alkyl or hydroxy-substituted lower alkyl), CH₂ Aa(wherein the Aa representation is the same as the above A representationexcluding hydroxy and amino) or --CH═N--R¹², R¹, R², R³, R⁴, W₁ and W₂are all hydrogen.

When X is CONHOH, R¹, R², R³ and R⁴ are all hydrogen. When X is loweralkoxycarbonyl, R⁴ is hydrogen, chlorine, carbamoyl, lower alkyl or--CH₂ CH₂ R^(7a) (wherein R^(7a) is bromine or di-lower alkylamino).

When X is aminomethyl, R¹, R² and R³ are hydrogen, and when W₁ and W₂are combined together to represent oxygen, R⁴ is hydrogen or amino andwhen W₁ and W₂ are hydrogen, R⁴ is hydrogen. When X and Y are combinedtogether to represent, aas --X--Y--, --O--, --CH₂ --O--, --CH₂--O--CO--O--, --CH₂ --O--CS--O--, --CH₂ --NR^(13a) --CO--O-- (whereinthe R^(13a) representation is the same as the above R¹³ representationexcluding hydrogen and lower alkyl), --CH₂ --NH--CS.sub.[O--, --CH₂--O--SO--O-- or ##STR11## R¹, R², R³, R⁴, W₁ and W₂ are all hydrogen.When X and Y are combined together to represent, as --X--Y--, --CH₂--NR^(13b) --CO--O-- (wherein R^(13b) is hydrogen or lower alkyl), R¹,R², R³ and R⁴ are all hydrogen. When R¹ is methyl, hydroxy,hydroxymethyl, lower alkoxy, bromine, chlorine or --NR⁵ R⁶, R² and R⁴are hydrogen. When R² is amino, R¹ and R⁴ are hydrogen. Compoundswherein R¹, R², R³, R⁴, W₁ and W₂ are hydrogen, X is methoxycarbonyl andY is hydroxy are excluded.

The compounds represented by general formula (I) are hereinafterreferred to as Compound (I). Compounds represented by formulae withother numbers are referred to similarly.

In the definitions of the groups in formula (I), the lower alkyl moietyin the lower alkoxy, the lower alkylaminocarbonyl, the lower alkyl, thedi-lower alkylamino, the lower alkoxycarbonyl, the hydroxy-substitutedlower alkylamino, the lower alkylthio, the lower alkylsulfenyl, thechlorine-substituted lower alkyl, the carboxylic acid-substituted loweralkyl and the lower alkoxycarbonyl-substituted lower alkyl includes astraight chain or branched alkyl having 1 to 5 carbon atoms, forexample, methyl, ethyl, n-propyl, i-propyl, n-butyl and n-pentyl.

As the hydroxy-substituted lower alkylamino in the definition of R⁷,2-hydroxyethylamino may be mentioned. As the hydroxy-substituted loweralkyl in the definitions of R⁸ and R⁹, 2-hydroxyethyl may be mentioned.In the definitions of R¹⁰ and R¹¹, the chlorine-substituted lower alkylincludes 2-chloroethyl. In the definitions of R¹⁰ and R¹¹, thecarboxylic acid-substituted lower alkyl includes hydroxycarbonylmethyl.In the definitions of R¹⁰ and R¹¹, the lower alkoxycarbonyl-substitutedlower alkyl includes lower alkoxycarbonylmethyl. In the definitions ofR¹⁰ and R¹¹, the dihydroxy-substituted lower alkyl includes2,3-dihydroxypropane.

In the definitions of R¹⁰ and R¹¹, the α-amino acid includes glycine,alanine, valine, proline, etc. in L-form, D-form and racemicmodification.

In cases where Compound (I) is aan acidic compound, base addition saltsthereof can be formed, and in cases where Compound (I) is a basiccompound, acid addition salts thereof can be formed. The base additionsalts of Compound (I) includes ammonium salts; alkali metal salts suchas lithium, sodium and potssium salts; alkaline earth metal salts suchas calcium and magnesium salts; salts with organic bases such astriethylamine, morpholine, piperidine and dicyclohexylamine; and saltswith basic amino acids such as arginine and lysine. The acid additionsalts of Compound (I) include hydrochloride, hydrobromide, sulfate,nitrate, formate, acetate, benzoate, maleate, fumarate, succinate,tartrate, citrate, oxalate, methanesulfonate, toluenesulfonate,aspartate, glutamate, etc. Non-toxic and pharmacologically acceptablesalts, for example, the base addition salts and acid addition saltsmentioned above are preferred, but other salts are also useful inisolating and purifying the product.

In Compound (I), the two carbon atoms to which CH₃ and X are bound areasymmetric carbons. The compounds in accordance with the presentinvention are obtained from optically active K-252 and KT-5556 throughreactions which do not alter steric nature and all of them are opticallyactive compounds having the same steric configuration as K-252 andKT-5556.

The process for producing Compound (I) is described below. Compound (I)can be produced by the following reaction steps.

Me, Et, Pr, Bu, Ph, Ac, Bzl, Py and Ts in the structural formulae,tables, etc. refer to methyl, ethyl, propyl, butyl, phenyl, acetyl,benzyl, pyridyl and toluenesulfonyl groups, respectively. ##STR12##

In the above formulae, R¹, R², R³, R⁸, R⁹, R¹², R¹³ and B have the samesignificance as described above. R¹⁵, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²⁵ and R²⁹are lower alkyl and R¹⁶ is hydrogen or lower alkyl. R²¹ is a residue ofan α-amino acid having the amino group protected with t-butoxycarbonylor benzyloxycarbonyl in which the hydroxy of the carboxylic acid isremoved, R^(21a) is a residue of an α-amino acid having the amino groupprotected with benzyloxycarbonyl in which the hydroxy of the carboxylicacid is removed; R^(21b) is a residue of an α-amino acid having theamino group protected with t-butoxycarbonyl in which the hydroxy of thecarboxylic acid is removed; and R²² is a residue of an α-amino acid inwhich the hydroxy of the carboxylic acid is removed.

One of R²³ and R²⁴ is hydrogen and the other is lower alkyl or allyl; orboth are lower alkyl or chlorine-substituted lower alkyl; or R²³ and R²⁴are combined together to represent --CH₂ CH₂ --B--CH₂ CH₂ --.

D is oxygen or sulfur. R²⁶ is lower alkyl or allyl. R²⁷ is guanidino.R²⁸ is lower alkyl or bromine-substituted lower alkyl. R³⁰ is amino,amino-substituted lower alkyl or hydroxy-substituted lower alkyl. R³¹ ishydroxy-substituted lower alkylamino. Xd is hydroxymethyl or loweralkoxycarbonyl. Z is halogen such as chlorine, bromine or iodine. E ischlorine or bromine. Compounds of formula Nos. (I-1), (I-2), . . . areincluded in Compound (I). Compounds of compound Nos. with suffixes a andb in the compound Nos. are inclined in the compounds of the same numberaccompanied by no suffix and compounds with ab is are included incompounds with a. For example, (I-1)a and (I-28)ab are included in (I-1)and (I-28)a, respectively.

Each step is described below.

Step 1

Compound (II) can be obtained by reaction of KT-5556 with aceticanhydride in a suitable solvent in the presence of a base. As the base,pyridine, N,N-dimethylaminopyridine, triethylamine, etc. can be used.Acetic anhydride and the base are usually used in an amount of 1.1 to 2equivalents based on KT-5556. As the reaction solvent, chloroform,dichloromethane and the like are used. The reaction is usually carriedout at 0° C. to room temperature and completed in 15 minutes to severalhours (Step 1-1).

Compound (II) is heated under reflux in thionyl chloride to give acidchloride of Compound (II), i.e., Compound (III) (Step 1-2). Compound(III) is made to react with an amine ##STR13## to obtain an amidecompound. The amine component is generally used in an equivalent orexcess amount, usually in an amount of 1 to 5 equivalents, based onCompound (III). As the reaction solvent, anhydrous chloroform,dichloromethane, and the like can be used. The reaction is usuallycarried out at room temperature and completed in several hours.

Then, the acetoxy group of the amide compound obtained above ishydrolyzed with an alkali, whereby Compound (I-1) can be obtained. Thereaction is carried out in a solvent mixture obtained by adding water toalcohols such as methanol and ethanol, such sodium hydroxide orpotassium hydroxide in an amount of 1.5 to 3 equivalents based on theamide compound. The reaction is usually carried out at room temperatureand completed in several hours (Step 1-3).

Step 2

Alcohol R₁₅ OH and an excess of thionyl chloride are added to KT-5556,and the mixture is heated under reflux to give compound (I-2). Thionylchloride is usually used in an amount of about one tenth (volume ratio)of the amount of the alcohol used also as the solvent. The reaction iscarried out in the range of 80° to 100° C., and almost completed inseveral hours to one day.

Step 3

Compound (I-2) and a suitable chlorinating agent, for example,N-chlorosuccinimide (NCS) are subjected to reaction in a solvent inertto the reaction to give Compound (I-3). The chlorinating agent isusually used in an equivalent amount based on Compound (I-2). The inertsolvent includes chloroform, dichloromethane, and the like. The reactionis carried out with heating under reflux, and usually completed inseveral hours.

Step 4

Compound (I-2) and halogen in an amount of 1 to 2 equivalents based onthe compound are stirred in an inert solvent at room temperature for oneday to give Compound (I-4). The inert solvent includes pyridine,chloroform, and the like.

Step 5

Compound (I-2) and a suitable carbamoylating agent, for example,chlorosulfonyl isocyanate are stirred in an inert solvent, for example,tetrahydrofuran (hereinafter referred to as THF) under ice cooling for 1to 3 hours, and then water is added thereto. The mixture is stirredunder heating at 70° to 80° C. for 0.5 to 1 hour to give Compound (I-5).The carbamoylating agent is used in an amount of 5 to 10 equivalents andwater is used in an large excess amount based on Compound (I-2).

Step 6

Compound (I-2) is allowed to react with acetic anhydride in the presenceof a base to give Compound (IV). The base includes pyridine,triethylamine, etc. Acetic anhydride is usually used in an amount of 5to 10 equivalents based on Compound (I-2). The reaction is usuallycarried out using pyridine as the solvent at room temperature andcompleted in 1 to several hours.

Step 7

Compounds (V-1), (V-2) and (V-3) are obtained by reaction of Compound(IV) with a suitable nitrating agent, for example, nitroniumtetrafluoroborte in a solvent inert to the reaction. The nitrating agentis usually used in an amount of 1 to 1.1 equivalents based on Compound(IV). The inert solvent includes sulfolane, acetonitrile, etc. Thereaction is carried out at room temperature to 80° C. and completedusually in 1 to 2 hours.

Step 8

Compound (VI-1) is obtined by reducing Compound (V-1) in a solvent inertto the reaction according to a suitable reduction method, for example,catalytic reduction. The catalyst includes 5 to 10% palladium carbon,etc. and is usually used in an amount of 0.1 to 0.5 times that of theweight of Compound (V-1). The inert solvent includes THF,dimethylformamide (hereinafter referred to as DMF), etc. The reaction isusually carried out at room temperature and completed in several hoursto one day.

Step 9

The reaction is carried out in a similar manner as in Step 8.

Step 10

The reaction is carried out in a similar manner as in Step 8.

Step 11

Compound (I-6) is obtained by reaction of Compound (VI-1) with sodiumlower alkoxide NaOR¹⁵ in an inert solvent, followed by acidificationwith hydrochloric acid, etc. NaOR¹⁵ is usually used in an amount of 5 to7 equivalents based on Compound (VI-1). The inert solvent includesdichloromethane, THF, etc. The reaction is carried out at 0° C. to roomtemperature and usually completed in 3 to 30 minutes.

Step 12

The reaction is carried out in a similar manner as in Step 11.

Step 13

The reaction is carried out in a similar manner as in Step 11.

Step 14

Compound (I-9) is obtained by reaction of Compound (I-6) with R¹⁶ CHOand a suitable reducing agent, for example, sodium cyanoborohydride in asolvent inert to the reaction. Usually, R¹⁶ CHO is used in a largeexcess amount and the reducing agent is used in an amount of 1 to 2equivalents based on Compound (I-6). As the inert solvent, a 1:1 solventmixture of THF and a suitable lower alkanol, for example, methanol, etc.can be used. The reaction is usually carried out at room temperature andcompleted in 0.5 to one hour.

Step 15

Compound (VII-1) is obtained by reaction of Compound (VI-1) with loweralkyl isocyanate R¹⁷ N═C═O in a solvent inert to the reaction in thepresence of a base. The base includes triethylamine, etc. Usually, R¹⁷N═C═O is used in an amount of 2 to 3 equivalents and the base is used inan amount of 1 to 2 equivalents based on Compound (VI-1). The inertsolvent includes dichloromethane, chloroform, etc. The reaction isusually carried out at room temperature and completed in 1 to 5 hours.

Step 16

Compound (VI-1) is allowed to react with potassium cyanate usually in anamount of about 5 equivalents based on the compound in a solvent mixtureof THF, acetic acid and water (10:1:1) to give Compound (VII-2). Thereaction is usually carried out at room temperature and completed in onehour.

Steps 17 and 18

The reactions are carried out in a similar manner as in Step 11.

Step 19

Compounds (VIII-1) and (VIII-2) are obtained by reaction of Compound(IV) with acetyl chloride in a solvent inert to the reaction in thepresence of an appropriate Lewis acid, for example, aluminum chloride.Usually, acetyl chloride is used in an amount of 1 equivalent and Lewisacid in an amount of 5 equivalents based on Compound (IV). The inertsolvent includes dichloromethane, chloroform, etc. The reaction isusually carried out under ice cooling and completed in several hours.

Step 20

Compound (IX-1) is obtained by reaction of Compound (VIII-1) with anappropriate oxidizing agent, for example, m-chloroperbenzoic acid in asolvent inert to the reaction, which is usually chloroform. Theoxidizing agent is usually used in an amount of 5 equivalents based onCompound (VIII-1) twice at an interval of one hour. The reaction isusually carried out with heating under reflux and completed in severalhours.

Step 21

The reaction is carried out in a similar manner as in Step 20.

Steps 22 and 23

The reactions are carried out in a similar manner as in Step 11.

Step 24

Compound (I-14) is obtained by reaction of Compound (I-12) with loweralkyl halide R¹⁸ Z in a solvent inert to the reaction in the presence ofa base. As the lower alkyl halide, iodides and bromides which are highlyreactive are preferred. The base includes sodium hydride, potassiumt-butoxide, etc. The lower alkyl halide and the base are usually used inan equivalent amount based on Compound (I-12). The inert solventincludes DMF, THF, etc. The reaction is usually carried out at 0° C. toordinary temperature and completed in 20 minutes to one hour.

Step 25

The reaction is carried out in a similar manner as in Step 24.

Step 26

Compound (X) is obtained by reaction of Compound (IV) withdichloromethyl methyl ether in a solvent inert to the reaction in thepresence of an appropriate Lewis acid, for example, titaniumtetrachloride. Usually, dichloromethyl methyl ether is used in an amountof 1 to 2 equivalents and titanium tetrachloride in an amount of 5 to 7equivalents based on Compound (IV). As the inert solvent,dichloromethane is usually used. The reaction is usually carried out atroom temperature and completed in several hours.

Step 27

Compound (XI) is obtained by reaction of Compound (X) with anappropriate reducing agent, for example, sodium borohydride in a solventinert to the reaction. The reducing agent is usually used in an amountof 2 to 3 equivalents based on Compound (X). As the inert solvent, asolvent mixture of chloroform-methanol (1:1) is usually used. Thereaction is usually carried out under ice cooling and completed in 0.5to one hour.

Step 28

Compound (XII) is obtained by reaction of Compound (XI) with loweralkylthiol R¹⁹ SH in a solvent inert to the reaction in the presence ofan appropriate acid catalyst, for example, camphor sulfonic acidcatalyst. Usually, R¹⁹ SH is used in an amount of 5 to 10 equivalentsand the acid is used in an equivalent amount based on Compound (XI). Theinert solvent includes chloroform, etc. The reaction is usually carriedout at room temperature and completed in 2 to 3 hours.

Step 29

Compound (XIII) is obtained by heating Compound (XII) under reflux withRaney nickel in an amount of 0.1 to 0.5 times the weight of Compound(XII) in ethyl acetate for 5 to 7 hours.

Steps 30 and 31

The reactions are carried out in a similar manner as in Step 11.

Step 32

Compound (I-18) can be obtained by reducing the ester moiety of Compound(I-2), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13),(I-14), (I-15), (I-16) or (I-17). As the reducing agent, lithiumaluminum hydride (1 to 2 molar equivalents based on the compoundsmentioned above) and sodium borohydride (5 equivalents based on thecompounds mentioned above) are useful. The reaction is carried out, inthe case of lithium aluminum hydride, in a solvent such as THF ordioxane, and in the case of sodium borohydride, in a solvent mixture ofTHF-MeOH, usually at 0° C. to room temperature and completed in severalhours.

Step 33

Tosylate (XIV) can be obtained by reaction of Compound (I-18)a withp-toluenesulfonyl chloride in the presence of a base. As the base,triethylamine, pyridine, N,N-dimethylaminopyridine, sodium hydride, etc.are used, and as the solvent, THF, dioxane, chloroform, etc. are used.p-Toluenesulfonyl chloride and the base are usually used in an amount of2 to 3 equivalents based on Compound (I-18)a. The reaction is usuallycarried out at 0° C. to room temperature and almost completed in severalhours to one day.

Step 34

Compound (I-19) is obtained by reaction of Compound (I-18)a withsuccinic anhydride in an inert solvent in the presence of an appropriatebase, for example, triethylamine at 80° to 110° C. for 2 hours.

Succinic anhydride is used in an amount of 1 to 2 equivalents and thebase in an amount of 1 to 2 equivalents based on Compound (I-18)a. Theinert solvent includes THF, DMF, etc.

Step 35

Compound (I-20) is obtained by heating Compound (I-18)a under refluxwith 3,4-dihydro-2H-pyran in an inert solvent for 4 hours in thepresence of an appropriate acid catalyst, for example, camphor sulfonicacid catalyst. 3,4-Dihydro-2H-pyran is used in an equivalent amount andthe acid in an amount of 0.1 equivalent based on the compound. The inertsolvent includes chloroform, THF, etc.

Step 36

The azide (I-21) can be obtained by reaction of Compound (XIV) withsodium azide (1 to 2 equivalents). As the solvent, DMF,dimethylsulfoxide, THF, etc. are used. The reaction is usually carriedout at room temperature and completed in several hours to one day.

Step 37

The amine (I-22) is obtained by reducing Compound (I-21) with lithiumaluminum hydride (excess, 2 to 6 molar equivalents). As the solvent,THF, dioxane, etc. are used. The reaction is usually carried out at 0°C. to room temperature and completed in several hours.

Step 38

Compound (I-23) can be obtained by stirring Compound (I-22) and abromoacetic acid lower alkyl ester in an inert solvent at roomtemperature for one day in the presence of an appropriate base, forexample, 4-dimethylaminopyridine. The bromoacetic acid lower alkyl esteris used in an amount of 1 to 2 equivalents and the base in an amount of2 equivalents based on Compound (I-22). The inert solvent includes DMF,THF, etc.

Step 39

Compound (I-24) can be obtained by adding 1N sodium hydroxide aqueoussolution to Compound (I-23), stirring the mixture in a solvent mixtureof THF and methanol (3:1) at room temperature for 0.5 hour, and thentreating the mixture with an acid. The 1N sodium hydroxide aqueoussolution is used in an amount of 3 to 5 equivalents based on Compound(I-23).

Step 40

Compound (I-25) can be obtained by stirring Compound (I-22) andN,N-dimethylformamide dimethyl acetal in an inert solvent at roomtemperature for 2 hours. N,N-Dimethylformamide dimethyl acetal is usedin an equivalent amount based on Compound (I-22). As the inert solvent,DMF is used.

Step 41

Compound (XV) can be obtained by condensing Compound (I-22) with R²¹ OHin a THF solvent using N-oxysuccinimide and dicyclohexyl carbodiimide(DCC). R²¹ OH, N-oxysuccinimide and DCC are used respectively in amountsof 1 to 2 equivalents, one equivalent and 1 to 2 equivalents based onCompound (I-22). The reaction is usually carried out at 0° C. to roomtemperature and completed in one day.

Step 42

The reaction is carried out in a similar manner as in Step 8.

Step 43

Compound (I-26) is obtained by reaction of Compound (XV)b with a largeexcess of trifluoroacetic acid in a chloroform solvent at roomtemperature for one hour.

Step 44

The epoxide (I-27) can be obtained by reaction of Compound (XIV) with anexcess (usually 1 to 2 equivalents) of sodium hydride. The reaction isusually carried out in THF or dioxane at room temperature and completedin several hours.

Step 45

The substituted amino compound (I-28) or the guanidino compound (I-29)can be obtained by reaction of Compound (XIV) with an amine component,namely, amine ##STR14## or guanidine in a large excess, usually in anamount of 1 to 10 equivalents. The reaction proceeds rapidly by adding astrong base such as 1,8-diazabicyclo[5,4,0]-7-undecene (DBU) in additionto the amine component which is allowed to react. In cases where theamine component used is a salt such as hydrochloride, it is necessary toadd an equimolar amount of tertiary amine such as triethylamine or DBU.The reaction is carried out in DMF or THF at room temperature andcompleted in several hours to one day.

Step 46

Compound (I-30) can be obtained by stirring Compound (I-28)ab and anequivalent amount of osmic acid in a solvent mixture of THF and pyridine(10:1) at room temperature for 0.5 hour, adding sodium hydrogensulfitein an amount of 4.4 equivalents thereto, and then stirring the mixtureat room temperature for one hour.

Step 47

Compound (I-31) can be obtained by reaction of Compound (XIV) withsodium thiolate (R²⁵ SNa) (excess, 1.5 to 10 equivalents). The reactionis usually carried out in THF at room temperature and completed inseveral hours to one day.

Step 48

The sulfoxide (I-32) is obtained by oxidizing Compound (I-31) withperiodic acid (1 to 1.05 equivalents). As the solvent, a solvent mixtureof THF and water is used. The reaction is usually carried out in therange of 0° C. to room temperature and completed in several hours to onenight.

Step 49

The oxo compound (I-33) is obtained by oxidizing Compound (I-18)a withperiodic acid (1 to 1.05 equivalents). The reaction is usually carriedout in a solvent mixture of methanol-THF at room temperature andcompleted in several hours.

Step 50

Compound (I-34) can be obtained by adding sodium borohydride in anamount of 5 equivalents to a solution of Compound (I-33) in aTHF-ethanol solvent mixture and stirring the mixture at room temperaturefor one hour.

Step 51

Compound (I-35) is obtained by reaction of Compound (I-2) with anappropriate reducing agent, for example, lithium aluminum hydride,usually in THF. The reducing agent is usually used in an equivalentamount. The reaction is carried out under ice cooling and usuallycompleted in one hour.

Step 52

Compound (I-36) is obtained by reaction of Compound (I-35) with H₂ NR¹²usually in a solvent mixture of THF-H₂ O (10:1). H₂ NR¹² is usually usedin an amount of 5 to 10 equivalents in the form of hydrochloride,hydrobromide or sulfate. The reaction is usually carried out at roomtemperature and completed in several hours to one day.

Step 53

Compound (I-37) is obtained by reaction of Compound (I-18)a with1,1'-carbonyldiimidazole or 1,1'-thiocarbonyldiimidazole in excess,usually in an amount of 1.5 to 3 equivalents. The reaction proceedswithout a base, but proceeds rapidly by the addition of a tertiary aminesuch as triethylamine. The reaction is usually carried out in THF atroom temperature and completed in several hours to one night.

Steps 54

The cyclic sulfinate (I-38) can be obtained by reaction of Compound(I-18)a with thionyl chloride in an amount of 1 to 1.5 equivalents inpyridine. The reaction is usually carried out at room temperature andcompleted in several hours.

Step 55

Compound (I-39) is obtained by reaction of Compound (I-22) withtrimethyl orthoacetate in an amount of 1 to 1.5 equivalents in sulfolaneat 100° C. for 3 hours in the presence of an appropriate acid catalyst,for example, camphor sulfonic acid catalyst (0.04 equivalent).

Step 56

The reaction is carried out in a similar manner as in Step 53.

Step 57

The reaction is caried out in a similar manner as in Step 53.

Step 58

The reaction is carried out in a similar manner as in Step 46.

Step 59

The reaction is carried out in a similar manner as in Step 49.

Step 60

The reaction is carried out in a similar manner as in Step 52.

Step 61

Compound (I-45) is obtained by reaction of Compound (I-40)a with loweralkyl halide R¹⁸ Z usually in DMF. As R¹⁸ Z, iodides which are highlyreactive are preferred. The reaction is usually carried out at roomtemperature and completed in several hours.

Step 62

Compound (I-46) is obtained by reaction of Compound (I-2) with anappropriate oxidizing agent, for example, Collins reagent (chromic aciddipyridine complex) in a pyridine solvent in the range of 0° C. to roomtemperature for one day. The oxidizing agent is used in an amount of 5to 7 equivalents based on Compound (I-2).

Step 63

Compound (XVI) is obtained by reaction of Compound (I-22) withbenzyloxycarbonyl chloride in a solvent mixture of THF-water under icecooling for 0.5 to one hour in the presence of an appropriate base, forexample, sodium hydrogencarbonate. The base is used in an amount of 5equivalents and benzyloxycarbonyl chloride in an amount of 1 to 1.5equivalents based on Compound (I-22).

Step 64

The reaction is carried out in a similar manner as in Step 62.

Step 65

The reaction is carried out in a similar manner as in Step 8.

Step 66

Compound (XIII) is obtained by heating Compound (I-18)a under refluxwith trimethyl orthoacetate in a chloroform solvent for 10-30 minutes inthe presence of an appropriate acid catalyst, for example, camphorsulfonic acid catalyst. The acid catalyst is used in an amount of 0.1equivalent and trimethyl orthoacetate in an amount of 2 equivalentsbased on Compound (I-18)a.

Step 67

The reaction is carried out in a similar manner as in Step 62.

Step 68

Compound (I-48) can be obtained by treating Compound (XIX) with a largeexcess of 3N hydrochloric acid aqueous solution in a chloroform-methanolsolvent mixture at room temperature for 1 to 2 hours and then treatingthe compound with a large excess of 2N sodium hydroxide aqueous solutionin a chloroform-methanol solvent mixture at room temperature for onehour.

Step 69

The reaction is carried out in a similar manner as in Step 32.

Step 70

Compound (XX) can be obtained by reaction of Compound (XIX) with R²⁸ Zin an amount of 1 to 10 equivalents in a DMF solvent under ice coolingin the presence of an appropriate base, for example, sodium hydride inan amount of 1 to 1.5 equivalents. The reaction is usually completed inseveral hours.

Step 71

The reaction is carried out in a similar manner as in Step 68.

Step 72

The reaction is carried out in a similar manner as in Step 70.

Step 73

The reaction is carried out in a similar manner as in Step 32.

Step 74

Compound (I-51) can be obtained by reaction of Compound (I-49)a orCompound (I-50)a with HNR₂ ²⁹ in an amount of 10 to 15 equivalents in aDMF solvent in the presence of an appropriate base, for example, DBU inan amount of 15 to 20 equivalents. The reaction is usually completed atroom temperature in one day.

Step 75

Compound (I-52) can be obtained by reaction of Compound (I-47) withhydrazine in an amount of 10 to 50 equivalents in dioxane at 70° to 110°C. for 4 to 10 hours.

Step 76

Compound (I-53) is obtained by reaction of Compound (I-48) with R³⁰ NH₂in an amount of 10 to 15 equivalents in an inert solvent at 70° to 110°C. for 4 to 10 hours. The reaction proceeds rapidly by using anappropriate base such as DBU. The inert solvent includes dioxane, DMF,etc.

Step 77

Compound (I-54) is obtained by reaction of Compound (I-53)a with R³¹ Zin an amount of 5 to 10 equivalents in a THF solvent in the presence ofan appropriate base, for example, triethylamine in an amount of 10equivalents. The reaction is usually completed at room temperature inone day.

Step 78

The reaction is carried out in a similar manner as in Step 62.

Step 79

Compound (I-56) is obtained by subjecting Compound (I-1)a to reactionwith exces acetic anhydride in a pyridine solvent at room temperaturefor several hours to protect the two hydroxy groups with acetyl,oxidizing the compound in a similar manner as in Step 66, and thentreating the compound with excess 28% aqueous ammonia in a DMF solventat room temperature for one day.

Isolation and purification of the product after completion of each stepdescribed above can be carried out by methods used in conventionalorganic synthesis, for example, by an appropriate combination ofextraction, crystallization, chromatography, etc.

Compound (I) shows a marked cell growth inhibitory activity againsthuman uterine cervical cancer Hela cells, human breast cancer cell MCF7, human colon adenocarcinoma cell COLO320DM and human lung differentedsquamous cell carcinoma cell PC-10, and accordingly, antitumorcompositions comprising Compound (I) as an effective ingredient areprovided.

Compound (I) and its pharmacologically acceptable salts includeolephilic and hydrophilic ones and such properties are particularlypreferred for pharmaceutical use in some cases. In cases where Compound(I) is used as an anti-tumor composition, each compound is dissolved inphysiological saline or a solution of glucose, lactose or mannitol forinjection, and usually intravenously administered as an injection in adose of 0.01 to 20 mg/kg. Alternatively, the compound may befreeze-dried in accordance with the Japanese Pharmacopoeia or may beprepared into injectable powder by adding sodium chloride thereto.Further, the anti-tumor composition may also contain pharmacologicallyacceptable well-known diluents, adjuvants and/or carriers such as saltswhich satisfy requirements for medical use. In cases where the compoundsis used as an injection, it is sometimes preferred to use auxiliaryagents which enhance the solubility. Doses may be appropriately varieddepending upon the age and conditions. Administration schedule can alsobe varied depending upon the conditions and dose. For example, thecompound is administered once a day (by single administration orconsecutive administration) or intermittently one to three times a weekor once every three weeks. Further, oral administration and rectaladministration are also possible in the same dose and in the samemanner.

The compound can be administered, with appropriate adjuvants, astablets, powders, granules, syrup, etc. for oral administration and assuppositories for rectal administration.

EXAMPLES

Representative examples of Compound (I) obtained by the processesdescribed above are shown in Table 2 and the intermediates thereof areshown in Table 3.

Examples of preparation of the Compound (I) and the intermediates arerespectively shown in Examples and Reference Examples. Compound Nos.correspond to Example Nos.

                                      TABLE 2                                     __________________________________________________________________________     ##STR15##                                  (I)                               Com-                                                                          pound                     W.sub.1                                             No. R.sup.1 R.sup.2                                                                          R.sup.3                                                                          R.sup.4 W.sub.2                                                                          X             Y                                  __________________________________________________________________________     1  H       H  H  H       H  CONHOH        OH                                  2  H       H  H  H       H  CONH.sub.2    OH                                  3  H       H  H  H       H  CONHCH.sub.2 CH.sub.2 OH                                                                    OH                                  4  H       H  H  H       H  CO.sub.2 Et   OH                                  5  H       H  H  Cl      H  CO.sub.2 Me   OH                                  6  Cl      H  H  H       H  CO.sub.2 Me   OH                                  7  Br      H  H  H       H  CO.sub.2 Me   OH                                  8  H       H  H  CONH.sub.2                                                                            H  CO.sub.2 Me   OH                                  9  NH.sub.2                                                                              H  H  H       H  CO.sub.2 Me   OH                                 10  NH.sub.2                                                                              H  NH.sub.2                                                                         H       H  CO.sub.2 Me   OH                                 11  H       NH.sub.2                                                                         H  H       H  CO.sub.2 Me   OH                                 12  NMe.sub.2                                                                             H  H  H       H  CO.sub.2 Me   OH                                 13  NHCONHMe                                                                              H  H  H       H  CO.sub.2 Me   OH                                 14  NHCONH.sub.2                                                                          H  H  H       H  CO.sub.2 Me   OH                                 15  OH      H  H  H       H  CO.sub.2 Me   OH                                 16  OH      H  OH H       H  CO.sub.2 Me   OH                                 17  On-Pr   H  H  H       H  CO.sub.2 Me   OH                                 18  CH.sub.2 OH                                                                           H  H  H       H  CO.sub.2 Me   OH                                 19  Me      H  H  H       H  CO.sub.2 Me   OH                                 20  H       H  H  H       H  CH.sub.2 OH   OH                                 21  OH      H  H  H       H  CH.sub.2 OH   OH                                 22  Cl      H  H  H       H  CH.sub.2 OH   OH                                 23  NH.sub.2                                                                              H  H  H       H  CH.sub.2 OH   OH                                 24  Br      H  H  H       H  CH.sub.2 OH   OH                                 25  OH      H  OH H       H  CH.sub.2 OH   OH                                 26  Me      H  H  H       H  CH.sub.2 OH   OH                                 27  H       H  H  H       H  CH.sub.2 OCOCH.sub.2 CH.sub.2 CO.sub.2                                                      OH                                  28(*)                                                                            H       H  H  H       H                                                                                 ##STR16##    OH                                 29  H       H  H  H       H  CH.sub.2 N.sub.3                                                                            OH                                 30  H       H  H  H       H  CH.sub.2 NH.sub.2                                                                           OH                                 31  H       H  H  H       H  CH.sub.2 NHCH.sub.2 CO.sub.2 Me                                                             OH                                 32  H       H  H  H       H  CH.sub.2 NHCH.sub.2 CO.sub.2 H                                                              OH                                 33  H       H  H  H       H  CH.sub.2 NCHNMe.sub.2                                                                       OH                                 34  H       H  H  H       H  CH.sub.2 NHCOCH.sub.2 NH.sub.2                                                              OH                                 35  H       H  H  H       H                                                                                 ##STR17##    OH                                 36  H       H  H  H       H  CH.sub.2 O                                       37  H       H  H  H       H  CH.sub.2 NHMe OH                                 38  H       H  H  H       H                                                                                 ##STR18##    OH                                 39  H       H  H  H       H                                                                                 ##STR19##    OH                                 40  H       H  H  H       H                                                                                 ##STR20##    OH                                 41  H       H  H  H       H                                                                                 ##STR21##    OH                                 42  H       H  H  H       H  CH.sub.2 NHCH.sub.2 CHCH.sub.2                                                              OH                                  43(*)                                                                            H       H  H  H       H  CH.sub.2 NHCH.sub.2 CH(OH)CH.sub.2 OH                                                       OH                                 44  H       H  H  H       H                                                                                 ##STR22##    OH                                 45  H       H  H  H       H  CH.sub.2 SMe  OH                                  46(*)                                                                            H       H  H  H       H  CH.sub.2 S(O)Me                                                                             OH                                 47  H       H  H  H       H  O                                                48  H       H  H  H       H  H             OH                                 49  H       H  H  H       H  CHO           OH                                 50  H       H  H  H       H  CHNOH         OH                                 51  H       H  H  H       H                                                                                 ##STR23##    OH                                 52  H       H  H  H       H                                                                                 ##STR24##    OH                                 53  H       H  H  H       H  CH.sub.2OCOO                                     54  H       H  H  H       H  CH.sub.2OCSO                                     55  H       H  H  H       H  CH.sub.2OSOO                                     56  H       H  H  H       H  CH.sub.2 NC(Me)O                                 57  H       H  H  H       H  CH.sub.2 NHCOO                                   58  H       H  H  H       H  CH.sub.2 NHCSO                                   59  H       H  H  H       H  CH.sub.2 NMeCOO                                  60  H       H  H  H       H  CH.sub.2 N(CH.sub.2 CHCH.sub.2)COO                61(*)                                                                            H       H  H  H       H                                                                                 ##STR25##                                       62  H       H  H  H       H                                                                                 ##STR26##                                       63  H       H  H  H       H                                                                                 ##STR27##                                       64  H       H  H  H       H  CH.sub.2 NCS(Me)O                                65  H       H  H  H       O  CO.sub.2 Me   OH                                 66  H       H  H  H       O  CH.sub.2 NH.sub.2                                                                           OH                                 67  H       H  H  H       O  CH.sub.2 OH   OH                                 68  H       H  H  CH.sub.2 CH.sub.2 Br                                                                  O  CO.sub.2 Me   OH                                 69  H       H  H  CH.sub.2 CH.sub.2 NMe.sub.2                                                           O  CO.sub.2 Me   OH                                 70  H       H  H  CH.sub.2 CH.sub.2 NMe.sub.2                                                           O  CH.sub.2 OH   OH                                 71  H       H  H  NH.sub.2                                                                              O  CH.sub.2 NH.sub.2                                                                           OH                                 72  H       H  H  NH.sub.2                                                                              O  CH.sub.2 OH   OH                                 73  H       H  H  CH.sub.2 CH.sub.2 NH.sub.2                                                            O  CH.sub.2 OH   OH                                 74  H       H  H  H       O  CH.sub.2 NHCOO                                   75  H       H  H  H       O  CH.sub.2 N(Me)CO O                               76  H       H  H  H       O  CONHOH        OH                                 __________________________________________________________________________     The compounds of Examples 9, 11, 12, 23, 30-32, 34, 35, 41, 42, 69-71 and     73 are hydrochlorides.                                                        (*)a mixture of diastereoisomers (about 1:1)                             

                                      TABLE 3                                     __________________________________________________________________________     ##STR28##                                                                        Refer-                                                                    Com-                                                                              ence                                                                      pound                                                                             Example           W.sub.1                                                 No. No.  R.sup.1                                                                            R.sup.2                                                                          R.sup.3                                                                          R.sup.4                                                                         W.sub.2                                                                          X           Y                                        __________________________________________________________________________    a   1    H    H  H  H H  CO.sub.2 H  OAc                                      b   2    H    H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      c   3    NH.sub.2                                                                           H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      d   3    NH.sub.2                                                                           H  NH.sub.2                                                                         Ac                                                                              H  CO.sub.2 Me OAc                                      e   3    H    NH.sub.2                                                                         H  Ac                                                                              H  CO.sub.2 Me OAc                                      f   4    Ac   H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      g   4    Ac   H  Ac Ac                                                                              H  CO.sub.2 Me OAc                                      h   5    CHO  H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      i   6    CH.sub.2 OH                                                                        H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      j   7    CH.sub.2 SEt                                                                       H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      k   8    Me   H  H  Ac                                                                              H  CO.sub.2 Me OAc                                      l   9    H    H  H  H H  CH.sub.2 OTs                                                                              OH                                       m   10   H    H  H  H H  CH.sub.2 NHCOCH.sub.2 NHCbz                                                               OH                                       n   11   H    H  H  H H                                                                                 ##STR29##  OH                                       o   12   H    H  H  H H  CH.sub.2 NHCbz                                                                            OH                                       p   13   H    H  H  H O.sub.2                                                                          CH.sub.2 NHCbz                                                                            OH                                       __________________________________________________________________________

EXAMPLE 1

A solution of 2.5 g of Compound a of Reference Example 1 in 60 ml ofthionyl chloride was heated under reflux for 2 hours. Thionyl chloridein the reaction solution was removed under reduced pressure and 40 ml ofethyl ether was added to the solid residue, followed by stirring.Insoluble matters were separated by filtration, washed with ethyl ether,and dried under reduced pressure to give 2.29 g (88%) of o-acetylatedacid chloride (III) as pale yellow powder.

To a solution of 206 mg (0.4 mmol) of Compound (III) in 5 ml ofanhydrous (P₂ O₅) chloroform were added 278 mg (4 mmol) of hydroxyaminehydrochloride and 0.56 ml (4 mmol) of triethylamine. After stirring atroom temperature for 6 hours, 1 ml of 1N sodium hydroxide aqueoussolution and 5 ml of methanol were added to the mixture, followed bystirring for one hour. To the reaction mixture was added 70 ml of THF,and the resulting solution was washed with 1N hydrochloride acid andsaturated sodium chloride aqueous solution, and then dried overanhydrous sodium sulfate. The solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography [eluent (hereinafter the same shall apply):chloroform-methanol] to give 91 mg (49%) of Compound 1 as pale yellowpowder.

Melting point: 259°-263° C. (CH₃ OH).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.28(d, 1H, J=8 Hz), 8.15-7.8(m, 2H),7.7-7.15(m, 5H), 7.06(dd, 1H, J=5, 7 Hz), 5.06(d, 1H, J=17 Hz), 4.86(d,1H, J=17 Hz), 3.36(dd, 1H, J=7, 14 Hz), 2.33(dd, 1H, J=5, 14 Hz),2.26(s, 3H).

MS (m/z): 469 (M⁺ +1).

EXAMPLE 2

Compound 2 (93 mg, 51%) was obtained as pale yellow powder from Compounda of Reference Example 1 and 28% aqueous ammonia in a similar manner asin Example 1.

Melting point: 262°-265° C. (CH₂ Cl₂ --CH₃ OH).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.30(d, 1H, J=8 Hz), 8.15-7.1(m, 7H),7.05(dd, 1H, J=5, 7 Hz), 4.99(br s, 2H), 3.33(dd, 1H, J=7, 14 Hz),2.39(dd, 1H, J=5, 14 Hz), 2.29(s, 3H).

MS (m/z): 453 M⁺ +1).

EXAMPLE 3

Compound 3 (118 mg, 59%) was obtained as pale yellow powder fromCompound a of Reference Example 1 and ethanolamine in a similar manneras in Example 1.

Melting point: 237°-239° C.

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.29(d, 1H, J=8 Hz), 8.2-7.8(m, 3H),7.7-7.15(m, 4H), 7.04(dd, 1H, J=5, 7 Hz), 4.98(br s, 2H), 3.9-3.45(m,4H), 3.31 (dd, 1H, J=7, 14 Hz), 2.29(dd, 1H, J=5, 14 Hz), 2.23(s, 3H).

MS (m/z): 497 (M⁺ +1).

EXAMPLE 4

To a suspension of 227 mg (0.5 mmol) of KT-5556 in 20 ml of ethanol wasadded 1 ml of thionyl chloride, and the mixture was heated under reflux.After two and four hours, 1 ml each of thionyl chloride was furtheradded to the mixture, and heating under reflux was carried out for 8hours in total. Volatile matters in the reaction mixture were removedunder reduced pressure. The residue was purified by silica gel columnchromatography (chloroform-methanol) to give 160 mg (66%) of Compound 4as pale yellow powder.

Melting point: 193°-195° C.

¹ H-NMR (DMSO-d₆) δ: 9.22(d, 1H, J=7.6 Hz), 8.1-7.85 (m, 3H),7.55-7.25(m, 4H), 7.11(dd, 1H, J=4.9, 7.3 Hz), 5.04(d, 1H, J=17.7 Hz),4.98(d, 1H, J=17.7 Hz), 4.40(m, 2H), 3.38(dd, 1H, J=7.3, 13.9 Hz),2.17(s, 3H), 2.02(dd, 1H, J=4.9, 13.9 Hz), 1.43(t, 3H, J=7.1 Hz).

MS (m/z): 481 (M⁺).

IR (KBr): 3430, 1730, 1675, 1635, 1590, 1460, 745 cm⁻¹.

EXAMPLE 5

In 20 ml of chloroform was dissolved 467 mg (1 mmol) of K-252, and 133mg (1 mmol) of N-chlorosuccinimide and 164 mg (1 mmol) ofazobisisobutyronitrile were added to the solution. The mixture washeated under reflux for 3 hours, and then the solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (chloroform) to give 229 mg (46%) of Compound 5 as paleyellow powder showing a melting point of 125° to 129° C.

NMR (CDCl₃) δ: 2.20(s, 3H), 2.68(dd, 1H, J=5, 14 Hz), 3.43(dd, 1H, J=7,14 Hz), 4.12(s, 3H), 4.88(d, 1H, J=15 Hz), 5.04(d, 1H, J=15 Hz),6.87(dd, 1H, J=5, 7 Hz), 7.24-7.64(m, 5H), 7.84-8.00(m, 2H), 9.00(d, 1H,J=8 Hz).

MS (m/e): 501 (M⁺).

EXAMPLE 6

In 100 ml of chloroform was dissolved 1.87 g (4 mmol) of K-252, and 4.0ml (4 mmol) of 0.98 M Cl₂ /AcOH was added to the solution. The mixturewas stirred overnight at room temperature. The precipitates wereseparated by filtration to give 0.41 g (20%) of Compound 6 as brownpowder. Further, the filtrate was concentrated under reduced pressureand the residue was purified by silica gel column chromatography (0.5%MeOH/CHCl₃) to give 0.41 g (20%) of Compound 6.

Melting point: 250°-257° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.08-2.36(m, 1H), 2.20(s, 3H), 3.40(dd, 1H, J=7,14 Hz), 3.98(s, 3H), 5.01 (s, 2H), 7.10(dd, 1H, J=5, 7 Hz), 7.28-8.12(m,6H), 9.31(d, 1H, J=2 Hz).

MS (m/e): 501 (M⁺).

EXAMPLE 7

In 3 ml of pyridine was dissolved 93 mg (0.2 mmol) of K-252. Under icecooling, 0.024 ml (0.48 mmol) of bromine was added to the solution,followed by stirring overnight. After completion of the reaction,tetrahydrofuran was added to the reaction solution. The mixture waswashed successively with 5% sodium thiosulfate aqueous solution andsaturated sodium chloride aqueous solution, and then dried overanhydrous magnesium sulfate. The solvent was distilled off under reducedpressure, and the residue was recrystallized from tetrahydrofuran andmethanol to give 70 mg (64%) of Compound 7 as yellowish brown powdershowing a melting point of 251° to 252° C.

NMR (CDCl₃ -DMSO-d₆) δ: 1.92-2.30(m, 1H), 2.20(s, 3H), 3.12-3.60(m, 1H),4.00(s, 3H), 5.04(s, 2H), 6.36(s, 1H), 7.04-7.24(m, 1H), 7.36-8.22(m,6H), 8.64(br. s, 1H), 9.48(br. s, 1H).

MS (m/e): 547 (M⁺).

EXAMPLE 8

In 5 ml of THF was dissolved 93 mg (0.2 mmol) of K-252, and 0.17 ml (2mmol) of chlorosulfonyl isocyanate was added to the solution under icecooling. The mixture was stirred at the same temperature for 2 hours.Then, 1 ml of water was added to the mixture, followed by stirring at70° C. for one hour. The reaction solution was washed successively withsaturated sodium bicarbonate aqueous solution and saturated sodiumchloride aqueous solution, and then dried over anhydrous magnesiumsulfate. The solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (2% MeOH/CHCl₃)to give 85 mg (77%) of Compound 8 as colorless powder showing a meltingpoint of 280° to 285° C.

NMR (DMSO-d₆) δ: 2.17(dd, 1H, J=5, 14 Hz), 2.18(s, 3H), 3.92(dd, 1H,J=7, 14 Hz), 3.94(s, 3H), 5.28(d, 1H, J=18 Hz), 5.34(d, 1H, J=18 Hz),7.22(dd, 1H, J=5, 7 Hz), 7.32(t, 1H, J=7 Hz), 7.42(t, 1H, J=7 Hz),7.50-7.58(m, 2H), 7.95-8.01(m, 3H), 9.06(d, 1H, J=8 Hz).

MS (m/e): 554 (M⁺ +1).

EXAMPLE 9

In 35 ml of dichloromethane was dissolved 700 mg (1.22 mmol) of Compoundc of Reference Example 3, and 1.2 ml (6.1 mmol) of 28% sodiummethylate/methanol solution was added to the solution. After fiveminutes, 3N hydrochloric acid aqueous solution was added to the mixture.The solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography(chloroform/methanol/DMF=80:10:10). Recrystallization fromchloroform-ether gave 507 mg (80%) of Compound 9 as yellow needlesshowing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.09(dd, 1H, J=5, 14 Hz), 2.18(s, 3H), 3.44(dd, 1H,J=7, 14 Hz), 3.96(s, 3H), 5.09 (s, 2H), 6.48(s, 1H), 7.24(dd, 1H, J=5, 7Hz), 7.18-7.71(m, 3H), 7.74-8.24(m, 3H), 8.77(s, 1H), 9.30(d, 1H, J=2Hz).

MS (m/e): 483 (MH⁺).

EXAMPLE 10

Compound 10 (53 mg, 41%) was obtained from 150 mg (0.26 mmol) ofCompound d of Reference Example 3 in a similar manner as in Example 9 ablack brown powder showing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 1.93(dd, 1H, J=5, 14 Hz), 2.10(s, 3H), 3.36(dd, 1H,J=7, 14 Hz), 3.94(s, 3H), 4.96 (br. s, 2H), 6.48-7.16(m, 3H), 7.24(d,1H, J=2 Hz), 7.64(d, 1H, J=2 Hz), 7.72(d, 1H, J=2 Hz), 8.62(d, 1H, J=2Hz).

MS (m/e): 498 (M⁺ +1).

EXAMPLE 11

Compound 11 (118 mg, 84%) was obtained from 150 mg (0.27 mmol) ofCompound e of Reference Example 3 in a similar manner as in Example 9 asblack brown powder showing a melting point higher than 300° C.

NMR (DMSO-d₆ +D₂ O) δ: 2.04(d, 1H, J=5, 14 Hz), 2.15 (s, 3H), 3.60(dd,1H, J=7, 14 Hz), 3.93(s, 3H), 5.00(d, 1H, J=18 Hz), 5.05(d, 1H, J=18Hz), 7.21-7.24(m, 1H), 7.27(t, 1H, J=8 Hz), 7.36-7.40(m, 2H),7.49-7.53(m, 1H), 7.96(d, 1H, J=8 Hz), 8.06 (d, 1H, J=7 Hz), 9.28(d, 1H,J=8 Hz).

MS (m/e): 483 (M⁺ +1).

EXAMPLE 12

In a solvent mixture of 3 ml of methanol and 3 ml of THF was dissolved155 mg (0.3 mmol) of Compound 9. To the solution were added 1 ml of 35%formaldehyde aqueous solution and then 0.3 mmol of sodiumcyanoborohydride. After the mixture was stirred at room temperature forone hour, 10% hydrochloric acid aqueous solution was added to adjust thepH to 1. The reaction mixture was washed with saturated sodium chlorideaqueous solution and dried over anhydrous magnesium sulfate. After thesolvent was distilled off under reduced pressure, the residue waspurified by silica gel column chromatography (5% methanol/chloroform)and recrystallized from chloroform-ethermethanol to give 50 mg (31%) ofCompound 12 as black brown powder showing a melting point higher than300° C.

NMR (DMSO-d₆) δ: 2.03(dd, 1H, J=5, 14 Hz), 2.16(s, 3H), 3.20-3.50(1H),3.40(6H), 3.93(s, 3H), 5.01 (d, 1H, J=17 Hz), 5.07(d, 1H, J=17 Hz),7.22(dd, 1H, J=5, 7 Hz), 7.36-7.53(m, 2H), 7.90-8.15(m, 4H), 8.75(s,1H), 9.44(s, 1H).

MS (m/e): 511 (M⁺).

EXAMPlE 13

In 10 ml of chloroform was dissolved 170 mg (0.3 mmol) of Compound c ofReference Example 3. To the solution were added 0.084 ml (0.6 mmol) oftriethylamine and then 0.88 ml (1.5 mmol) of methyl isocyanate. Themixture was stirred at room temperature for one hour, and 2 ml ofmethanol was added to the reaction mixture. The solvent was distilledoff under reduced pressure, and the residue was triturated with methanolto give 150 mg (80.2%) of Compound (VII-1, R¹⁵ =Me, R¹⁷ =Me0 as paleyellow powder showing a melting point higher than 300° C.

MS (m/e): 593 (M⁺ --NHMe).

Compound 13 (89 mg, 93.7%) was obtained from 10 mg (0.17 mmol) of theabove compound (VII-1, R¹⁵ =Me, R¹⁷ =Me) in a similar manner as inExample 9 as pale yellow powder showing a melting point higher than 300°C. (recrystallized from methanol).

NMR (CDCl₃ +DMSO-d₆) δ: 2.21(s, 3H), 2.28(dd, 1H, J=5, 14 Hz), 2.83(s,3H), 4.05(s, 3H), 4.96(br. s, 2H), 6.93(dd, 1H, J=5, 7 Hz), 7.28-7.64(m,3H), 7.84-8.04(m, 3H), 8.84(d, 1H, J=2 Hz).

MS (m/e): 509 (M⁺ --NHMe).

EXAMPLE 14

In a solvent mixture of 10 ml of THF and 1 ml of acetic acid wasdissolved 170 mg (0.3 mmol) of Compound c of Reference Example 3. To thesolution was added 1 ml of an aqueous solution of 120 mg (1.5 mmol) ofpotassium cyanate, and the mixture was stirred at room temperature forone hour. The solvent was distilled off under reduced pressure, and theresidue was triturated with water to give 178 mg (97.3%) of Compound(VII-1, R¹⁵ =Me) as yellow powder showing a melting point higher than300° C.

MS (m/e): 593 (M⁺ --NH₂).

Compound 14 (34 mg, 50%) was obtained from 80 mg (0.13 mmol) of theabove compound in a similar manner as in Example 9 as pale yellow powdershowing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.11(dd, 1H,, J=5, 14 Hz), 2.17(s, 3H), 3.20-3.63(1H),3.97(s, 3H), 5.79(br. s, 2H), 6.40(s, 1H), 6.97-7.23(m, 1H),7.30-7.70(m, 2H), 7.76-8.10(m, 4H), 8.70(s, 1H), 8.79(s, 1H), 9.20(s,1H), 9.30(s, 1H).

MS (m/e): 508 (M⁺ --NH₃).

EXAMPLE 15 In chloroform was dissolved 20 mg (0.033 mmol) of Compound fof Reference Example 4. To the solution was added 25 mg (0.15 mmol) ofm-chloroperbenzoic acid twice at an interval of one hour, followed byheating under reflux for 3 hours. After washing with saturated sodiumbicarbonate aqueous solution and water, the mixture was dried overanhydrous magnesium sulfate. The solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (chloroform) and recrystallized from chloroform-ether togive 10 mg (48.0%) of Compound (IX-1, R¹⁵ =Me) as brown powder showing amelting point higher than 300° C.

NMR (CDCl₃) δ: 1.79(s, 3H), 2.09(dd, 1H, J=5, 14 Hz), 2.26(s, 3H),2.40(s, 3H), 2.70(s, 3H), 3.94(dd, 1H, J=7, 14 Hz), 4.00(s, 3H), 5.34(s,2H), 6.98 (dd, 1H, J=5, 7 Hz), 7.20-7.70(m, 3H), 7.92-8.20(m, 3H),8.90(d, 1H, J=2 Hz).

MS (m/e): 610 (M⁺ +1).

Compound 15 (0.3 g, 38.8%) was obtained from 1.0 g (1.6 mmol of theabove compound in a similar manner as in Example 9 as reddish brownprisms showing a melting point higher than 300+ C. (recrystallized fromchloroform).

NMR (DMSO-d₆) δ: 1.97(dd, 1H, J=5, 14 Hz), 2.12(s, 3H), 3.35(dd, 1H,J=7, 14 Hz), 3.92(s, 3H), 5.01 (s, 2H), 6.32(s, 1H), 6.88-7.16(m, 2H),7.28-7.64(m, 2H), 7.72(d, 1H, J=8 Hz), 7.80-8.20(m, 2H), 8.60(s, 1H),8.71(d, 1H, J=2 Hz), 9.10(s, 1H).

MS (m/e): 484 (M⁺ +1).

EXAMPLE 16

Compound (IX-2, R¹⁵ =Me) (80 mg, 42%) was obtained as brown powder from182 mg (0.3 mmol) of Compound g of Reference Example 4 in a similarmanner as in Example 15.

NMR (CDCl₃) δ: 1.84(s, 3H), 1.96-2.40(m, 1H), 4.02(s, 3H), 5.36(s, 2H),6.72-7.08(m, 1H), 7.24-7.64 (m, 3H), 7.76-8.08(m, 2H), 8.48(s, 2H),9.01(d, 1H, J=2 Hz).

Compound 16 (10 mg, 15%) was obtained from 80 mg (0.13 mmol) of theabove compound in a similar manner as in Example 9 as yellow powdershowing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 1.97(dd, 1H, J=5, 14 Hz , 2.10(s, 3H), 3.00-3.50(1H),3.92(s, 3H), 4.94(s, 2H), 6.23(s, 1H), 6.80-7.12(m, 3H), 7.35(d, 1H, J=2Hz), 7.65(d, 1H, J=8 Hz), 7.76(d, 1H, J=8 Hz), 8.46(s, 1H), 8.67(d, 1H,J=2 Hz), 9.03(s, 1H), 9.20(s, 1H).

MS (m/e): 500 (M⁺ +1).

EXAMPLE 17

In 0.5 ml of DMF was suspended 9.6 mg (0.2 mmol) of 50% sodium hydride,and 1 ml of DMF solution of 96 mg (0.2 mmol) of Compound 15 was added tothe suspension under ice cooling. After ten minutes, 0.02 ml (0.2 mmol)of n-propyl iodide was added to the mixture at the same temperature,followed by stirring for one hour. After completion of the reaction,saturated ammonium chloride solution was added. The mixture wasextracted with chloroform, washed with saturated sodium chloride aqueoussolution, and dried over anhydrous magnesium sulfate. The solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (CHCl₃) and recrystallized fromdichloromethane-methanol to give 60 mg (57.1%) of Compound 17 as brownneedles showing a melting point of 228° to 230° C. (recrystallized fromchloroform).

NMR (DMSO-d₆) δ: 1.07(t, 3H, J=8 Hz), 1.72-2.24(m, 3H), 2.16(s, 3H),2.90-3.40(1H), 3.94(s, 3H), 4.08(t, 2H, J=7 Hz), 5.04(br.s, 2H), 6.34(s,1H), 7.00-7.24(m, 2H), 7.32-7.60(m, 2H), 7.76-8.16(m, 3H), 8.60(s, 1H),8.87(d, 1H, J=2 Hz).

MS (m/e): 526 (M⁺ +1).

EXAMPLE 18

Compound 18 (243 mg, 26%) was obtained from 1.11 g (1.91 mmol) ofCompound i of Reference Example 6 in a similar manner as in Example 9 asbrown powder showing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.00(dd, 1H, J=5, 14 Hz), 2.14(s, 3H), 3.26-3.43(m,1H), 3.92(s, 3H), 4.66(s, 2H), 4.97(d, 1H, J=18 Hz), 5.03(d, 1H, J=18Hz), 6.33 (s, 1H), 7.11(dd, 1H, J=5, 7 Hz), 7.33-7.37(m, 1H),7.45-7.50(m, 2H), 7.83(d, 1H, J=8 Hz), 7.49 (d, 1H, J=8 Hz), 8.60 (s,1H), 9.14(s, 1H).

MS (m/e): 497 (M⁺).

EXAMPLE 19

Compound 19 (70 mg, 82%) was obtained from 100 mg (0.17 mmol) ofCompound k of Reference Example 8 in a similar manner as in Example 9 aslight yellow powder showing a melting point higher than 300° C.

NMR (CDCl₃) δ: 2.12(s, 3H), 2.38(s, 3H), 2.95(dd, 1H, J=5, 14 Hz),3.48(dd, 1H, J=7, 14 Hz), 4.04 (s, 3H), 4.24(d, 1H, J=18 Hz), 4.48(d,1H, J=18 Hz), 5.42(s, 1H), 5.75(s, 1H), 6.78(dd, 1H, J=5, 7 Hz),6.94-7.20 (m, 2H), 7.28-7.62(m, 2H), 7.81(dd, 1H, J=2, 8 Hz), 8.00(d,1H, J=8 Hz), 8.40(s, 1H).

MS (m/e): 481 (M⁺).

EXAMPLE 20

A solution of 7.01 g (15 mmol) of K-252 in 100 ml of anhydrous THF wasice cooled, and 1.14 g (30 mmol) of lithium aluminum hydride was addedthereto, followed by stirring at room temperature for 2 hours. Aftermethanol was added to decompose excess reducing agent, the reactionmixture was filtered through Celite. The filtrate was washed with 1Nhydrochloric acid and saturated sodium chloride aqueous solution anddried over anhydrous sodium sulfate. The solvent was removed underreduced pressure, and the residue was purified by silica gel columnchromatography (chloroform-methanol) to give 5.34 g (81%) of Compound 20as pale yellow powder.

Melting point: 266°-275° C. (recrystallized from CH₃ OH).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.24(d, 1H, J=8 Hz), 8.2-7.7(m, 3H),7.6-7.0(m, 4H), 6.74(dd, 1H, J=5, 7 Hz), 4.90(d, 1H, J=18 Hz), 4.69(d,1H, J=18 Hz), 4.13(d, 1H, J=11 Hz), 3.91(d, 1H, J=11 Hz), 3.29 (dd, 1H,J=7, 14 Hz), 2.38(dd, 1H, J=5, 14 Hz), 2.19(s, 3H).

MS (m/z): 440 (M⁺ +1).

EXAMPLE 21

In 1 ml of THF was dissolved 48 mg (0.1 mmol) of Compound 15, and 38 ml(1 mmol) of lithium aluminum hydride was added to the solution, followedby stirring at room temperature for one hour. After 0.04 ml of water,0.04 ml of 15% sodium hydroxide aqueous solution and 0.04 ml of waterwere added successively, the reaction solution was extracted with THF.The extract was washed with saturated sodium chloride aqueous solutionand dried over anhydrous magnesium sulfate. The solvent was distilledoff under reduced pressure, and the residue was purified by silica gelcolumn chromatography (2% MeOH--CHCl₃) to give 15 mg (33%) of Compound21 as brown powder showing a melting point higher than 300° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.00-2.30(m, 1H), 2.20(s, 1H), 3.08-3.36(m, 1H),3.84-4.04(m, 2H), 4.52 (m, 1H), 4.96(s, 2H), 6.69(dd, 1H, J=5, 7 Hz),6.92-8.02(m, 6H), 8.63(s, 1H), 8.77(d, 1H, J=2 Hz).

MS (m/e): 455 (M⁺).

EXAMPLE 22

In a mixture of 15 ml of THF and 3 ml of methanol wad dissolved 412 mg(0.82 mmol) of Compound 6, and 218 mg (5.75 mmol) of sodium borohydridewas added to the solution under ice cooling, followed by stirring at thesame temperature for 3 hours. After 3N hydrochloric acid aqueoussolution was added, the reaction solution was extracted with THF. Theextract was washed with saturated sodium chloride aqueous solution anddried over anhydrous magnesium sulfate. The solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (3% MeOH--CHCl₃) to give 174 mg (45%) of Compound22 as brown powder showing a melting point of 275° to 280° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.08-2.28(m, 1H), 2.20(s, 3H), 3.12-3.36(m, 1H),3.76-4.00(m, 2H), 5.00 (s, 2H), 6.80-7.00(m, 1H), 7.20-8.16(m, 6H),8.40(br.s, 1H), 9.24(s, 1H).

MS (m/e): 474 (M⁺ +1).

EXAMPLE 23

Compound 23 (102 mg, 100%) was obtained from 110 mg (0.2 mmol) ofCompound 9 in a similar manner as in Example 22 as brown powder showinga melting point higher than 300° C.

NMR (DMSO-d₆ +D₂ O) δ: 1.80-2.10(m, 1H), 2.10(s, 3H), 3.04-3.20(m, 1H),3.84(br.s, 2H), 5.04(br.s, 2H), 6.92-7.16(m, 1H), 7.20-8.20(m, 7H), 9.23(d, 1H, J=0.2 Hz).

MS (m/e): 455 (MH⁺).

EXAMPLE 24

Compound 24 (22 mg, 26%) was obtained from 90 mg (0.16 mmol) of Compound7 in a similar manner as in Example 21 as brown powder showing a meltingpoint of 258° to 268° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.10(dd, 1H, J=5, 14 Hz), 2.20(s, 3H), 3.20(dd,1H, J=18, 14 Hz), 3.90(br.s, 2H), 5.00(s, 2H), 5.34(s, 1H), 6.88(dd, 1H,J=5, 8 Hz), 7.20-8.20(m, 6H), 8.36(s, 1H), 9.40(s, 1H).

MS (m/e): 518 (M⁺ +1).

EXAMPLE 25

Compound 25 (78 mg, 33%) was obtained from 250 mg (0.49 mmol) ofCompound 6 in a similar manner as in Example 21 as brown powder showinga melting point higher than 300° C.

NMR (DMSO-d₆ +D₂ O) δ: 2.00(dd, 1H, J=5, 14 Hz), 2.06 (s, 3H), 3.08(dd,1H, J=7, 14 Hz), 3.78(d, 1H, J=11 Hz), 3.97(d, 1H, J=11 Hz), 4.75(s,2H), 6.63 (dd, 1H, J=5, 7 Hz), 6.98(dd, 2H, J=2, 8 Hz), 7.26 (s, 1H),7.31(d, 1H, J=8 Hz), 7.70(d, 1H, J=8 Hz), 8.46(d, 1H, J=2 Hz).

MS (m/e): 472 (M⁺ +1).

EXAMPLE 26

Compound 26 (33 mg, 100%) was obtained from 35 mg (0.072 mmol) ofCompound 19 in a similar manner as in Example 22 as brown powder showinga melting point of 245° to 250° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.05(dd, 1H, J=5, 14 Hz), 2.20 (s, 3H), 2.54(s,3H), 3.16(dd, 1H, J=7, 14 Hz), 3.88(s, 2H), 4.98(s, 2H), 6.82(dd, 1H,J=5, 7 Hz), 7.20-7.60(m, 4H), 7.98(d, 1H, J=8 Hz), 9.06(s, 1H).

MS (m/e): 454 (M⁺ +1).

EXAMPLE 27

In 2 ml of DMF was dissolved in 44 mg (0.1 mmol) of Compound 20, and 22mg (0.22 mol) of succinic anhydride was added to the solution, followedby stirring at 100° C. for 2 hours. Then, 10 ml of water was added tothe reaction solution. The precipitates were separated by filtration andpurified by thin layer chromatography to give 25 mg (46.4%) of Compound27 as colorless powder showing a melting point of 198° to 208° C.

NMR (DMSO-d₆) δ: 2.09(dd, 1H, J=5, 14 Hz), 2.18(s, 3H), 2.61(t, 2H, J=6Hz), 2.70-2.74(m, 2H), 3.07(dd, 1H, J=7, 14 Hz), 4.38(d, 1H, J=11 Hz),4.50(d, 1H, J=12 Hz), 4.97(d, 1H, J=18 Hz), 5.03(d, 1H, J=17 Hz),5.83(s, 1H), 7.04(dd, 1H, J=5, 7 Hz), 7.27 (t, 1H, J=7 Hz), 7.34(t, 1H,J=7 Hz), 7.45-7.49 (m, 2H), 7.79(d, 1H, J=8 Hz), 8.00(d, 1H, J=8 Hz),8.05(d, 1H, J=8 Hz), 8.6(1H, s), 9.20(d, 1H, J=8 Hz), 12.32 (br.s, 1H).

MS (m/e): 540 (M⁺ +1).

EXAMPLE 28

In 10 ml of CHCl₃ was dissolved 88 mg (0.2 mmol) of Compound 21, and18.2 μml (0.2 mmol) of dihydropyran and 5 L of camphor sulfonate wereadded to the solution, followed by heating under reflux for 4 hours. Thereaction solution was washed successively with saturated sodiumbicarbonate aqueous solution and saturated sodium chloride aqueoussolution, and then dried over anhydrous magnesium sulfate. The solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (CHCl₃) to give 30 mg (29%) ofCompound 28 as colorless powder showing a melting point of 201° to 208°C.

NMR (CDCl₃) δ: 1.40-2.00(m, 6H), 2.20(s, 3H), 2.40-2.76(m, 1H),3.00-3.40(m, 1H), 3.52-4.28(m, 4H), 4.52-4.92(m, 3H), 5.50-6.30(m, 1H),6.50-6.80 (m, 1H), 7.08-7.64(m, 5H), 7.80-8.04(m, 2H), 8.99, 9.07(two d,1H, J=8 Hz).

MS (m/e): 524 (M⁺ +1).

EXAMPLE 29

A solution of 594 mg (1.0 mmol) of Compound l of Reference Example 9 and130 mg (2.0mmol) of sodium azide in 6 ml of DMF was stirred at roomtemperature overnight. To the reaction mixture was added 50 ml of THF,and the resulting solution was washed with an acid and an alkali. Thesolvent was removed under reduced pressure, and the residue was purifiedby silica gel column chromatography (chloroform-methanol) to give 405 mg(87%) of Compound 29 as pale yellow powder.

Melting point: 218°-223° C. (THF--CH₃ OH).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.31(d, 1H, J=8 Hz), 8.15-7.2(m, 7H),6.87(dd, 1H, J=5, 7 Hz), 5.00(s, 2H), 3.99(d, 1H, J=13 Hz), 3.56(d, 1H,J=13 Hz), 3.21 (dd, 1H, J=7, 14 Hz), 2.37(dd, 1H, J=5, 14 Hz), 2.9(s,3H).

MS (m/z): 465 (M⁺ +1).

IR (KBr): 3430, 2100, 1670, 1640, 1590, 1460, 745 cm⁻¹.

EXAMPLE 30

To a solution of 232 mg (0.5 mmol) of Compound 29 in 7 ml of anhydrousTHF was added 114 mg (3.0 mmol) of lithium aluminum hydride, followed bystirring at room temperature for 2 hours. To the reaction mixture wasadded 30 ml of THF, and the mixture was filtered through Celite. Thefiltrate was washed with an acid and an alkali. The solvent was removedunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (chloroform-methanol) to give 68 mg (31%) ofCompound 30 as pale yellow powder.

Melting point: >300° C. (CH₃ OH).

¹ H-NMR (DMSO-d₆) δ: 9.21(d, 1H, J=7.9 Hz), 8.1-7.7 (m, 3H),7.55-7.25(m, 4H), 7.00(dd, 1H, J=5.2, 7.4 Hz), 5.04(d, 1H, J=17.5 Hz),4.97(d, 1H, J=17.5 Hz), 3.25(dd, 1H, J=7.4, 13.6 Hz), 3.13(d, 1H, J=12.9Hz), 2.88(d, 1H, J=12.9 Hz), 2.12(s, 3H), 1.91(dd, 1H, J=5.2, 13.6 Hz).

MS (m/z): 439 (M⁺ +1).

IR (KBr): 3440, 1665, 1640, 1590, 745 cm⁻¹.

EXAMPLE 31

In 10 ml of DMF was dissolved 280 mg (0.64 mmol) of the free amine ofCompound 30, and 0.1 ml (1.05 mmol) of methyl bromoacetate and 156 ml(1.28 mmol) of 4-dimethylaminopyridine (DMAP) were added to thesolution, followed by stirring at room temperaturee for one day. Afterthe solvent was distilled off under reduced pressure, the residue waspurified by silica gel column chromatography (3% MeOH--CHCl₃) to give180 mg (55%) of Compound 31 as pale yellow powder showing a meltingpoint higher than 300° C.

NMR (DMSO-d₆) δ: 2.04-3.12(m, 1H), 2.18(s, 3H), 3.32-3.76(m, 1H),3.86(s, 3H), 4.23(br.s, 2H), 5.06(br.s, 2H), 6.42(br.s, 1H),7.00-7.80(m, 6H), 7.92-8.20(m, 2H), 8.69(br.s, 1H), 9.26 (d, 1H, J=8Hz).

MS (m/e): 511 (MH⁺).

EXAMPLE 32

In a mixture of 1 ml of THF and 0.3 ml of methanol was dissolved 15 mg(0.029 mmol) of the free amine of Compound 31, and 0.1 ml of 1N sodiumhydroxide aqueous solution was added to the solution, followed bystirring at room temperature for 0.5 hour. After the pH was adjusted to1 to 2 with 3N hydrochloric acid aqueous solution, the mixture wasconcentrated and the precipitates were separated by filtration. Theprecipitates were then washed with water to give 14 mg (93%) of Compound32 as brown powder showing a melting point higher than 300° C.

NMR (DMSO-d₆ +D₂ O)δ: 1.96-2.32(m, 1H), 2.15(s, 3H), 3.04-3.80(m, 3H),3.64(s, 2H), 5.00(br.s, 2H), 6.96-7.84(m, 5H), 7.92-8.20(m, 2H), 9.21(d,1H, J=8 Hz).

MS (m/e): 497 (MH⁺).

EXAMPLE 33

In 4 ml of DMF wad dissolved 438 mg (1 mmol) of the free amine ofCompound 30, and 0.16 ml (1.2 mmol) of N,N-dimethylformamidedimethylacetal was added to the solution, followed by stirring at roomtemperature for 2 hours. After the solvent was distilled off underreduced pressure, the residue was crystallized from 10%methanol-chloroform to give 371 mg (75%) of Compound 33 as brown powdershowing a melting point of 265° to 267° C.

NMR (DMSO-d₆) δ: 1.93(dd, 1H, J=5, 14 Hz), 2.17(s, 3H), 2.88(s, 6H),3.26(dd, 1H, J=7, 14 Hz), 3.64(s, 2H), 5.02(s, 2H), 5.12(s, 1H),7.00(dd, 1H, J=5, 7 Hz), 7.20-7.60(m, 4H), 7.58s, 1H), 7.84(d, 1H, J=8Hz), 8.04(t, 2H, J=8 Hz), 8.57(s, 1H), 9.24(d, 1H, J=8 Hz).

MS (m/e): 494 (M⁺ +1).

EXAMPLE 34

In 3 ml of DMF was dissolved 100 mg (0.15 mmol) of Compound m ofReference Example 10, and 100 mg of 10% palladium carbon was added tothe solution, followed by stirring at 40° C. for 5 hours in a stream ofhydrogen. The reaction mixture was filtered through Celite and thesolvent in the filtrate was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (MeOH--CHCl₃-28% NH₄ OH=5:95:0.25) and then dissolved in THF. 1.7N HCl/AcOEt wasadded to the solution to convert the product into the hydrochloride,whereby 40 mg (47%) of Compound 34 was obtained as pale yellow powdershowing a melting point higher than 300° C.

NMR (DMSO-d₆ +D₂ O) δ: 1.88-2.12(m, 1H), 2.18(s, 3H), 2.91(dd, 1H, J=7,14 Hz), 3.28(s, 2H), 3.72(br.s, 2H), 5.03(s, 2H), 6.96(dd, 1H, J=5, 7Hz), 7.20-7.64(m, 4H), 7.76(dd, 1H, J=2, 8 Hz), 7.90-8.16(m, 2H),9.16(d, 1H, J=8 Hz).

MS (m/e): 496 (MH⁺).

EXAMPLE 35

In a mixture of 5 ml of chloroform and 5 ml of trifluoroacetic acid wasdissolved 250 ml of Compound n of Reference Example 11. The solution wasstirred at room temperature for one hour, and the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (MeOH--CHCl₃ -28% NH₄ OH=5:95:0.25) and thendissolved in THF. 1.7N HCl/AcOEt was added to the solution to convertthe product into the hydrochloride, whereby 110 mg (49%) of Compound 35was obtained as pale yellow powder showing a melting point higher than300° C.

NMR (DMSO-d₆ +D₂ O) δ: 1.60-2.60(m, 5H), 2.20(s, 3H), 2.84-3.12(m, 1H),3.12-3.96(m, 4H), 4.28-4.52 (m, 1H), 5.03(s, 2H), 7.02(dd, 1H, J=5, 7Hz), 7.20-7.64(m, 4H), 7.80(d, 1H, J=8 Hz), 7.96-8.16 (m, 2H), 9.20(d,1H, J=8 Hz).

MS (m/e): 536 (MH⁺).

EXAMPLE 36

A solution of 1700 mg (2.9 mmol) of Compound l of Reference Example 9 in50 ml of anhydrous THF was ice cooled and 228 mg (5.8 mmol) of 60% oilsodium hydride was added thereto, followed by stirring at roomtemperature for 2.5 hours. The reaction solution was washed with an acidand an alkali. The solvent was removed under reduced pressure, and theresidue was purified by silica gel column chromatography(chloroform-methanol) to give 884 mg (73%) of Compound 36 as pale yellowpowder.

Melting point: 292°-296° C. (decomposed).

¹ H-NMR (DMSO-d₆) δ: 9.31(d, 1H, J=7.5 Hz), 8.1-7.75 (m, 3H),7.55-7.3(m, 4H), 7.22(dd, 1H, J=1.0, 6.0 Hz), 5.00(s, 2H), ca. 3.35(dd,1H), 3.29(d, 1H, J=4.4 Hz), 3.03(d, 1H, J=4.4 Hz), 2.446(s, 3H),2.00(dd, 1H, J=1.0, 14.7 Hz).

MS (m/z): 421 (M⁺).

IR (KBr): 3450, 1680, 1635, 1590, 1460, 1355, 1315, 1225, 750 cm⁻¹.

EXAMPLE 37

To a DMF solution of 126 mg (0.3 mmol) of Compound l of ReferenceExample 9 were added 202 mg (3.0 mmol) of methylamine hydrochloride and0.449 ml (3.0 mmol) of 1,8-diazabicyclo-[5,4,0]-7-undecene (DBU),followed by stirring at room temperature overnight. To the reactionmixture was added 50 ml of THF, and the resulting solution was washedwith saturated sodium chloride aqueous solution and then dried overanhydrous sodium sulfate. The solvent was removed under reduced pressureand the residue was purified by silica gel column chromatography(chloroformmethanol) to give 65 mg (48%) of Compound 37 as pale yellowamorphous powder.

¹ H-NMR (DMSO-d₆) δ: 9.21(d, 1H, J=7.9 Hz), 8.1-7.75 (m, 3H),7.5-7.25(m, 4H), 7.02(dd, 1H, J=5.3, 7.4 Hz), 5.03(d, 1H, J=17.6 Hz),4.96(d, 1H, J=17.6 Hz), 3.34(dd, 1H, J=7.4, 13.4 Hz), 3.07(d, 1H, J=11.7Hz), 2.86(d, 1H, J=11.7 Hz), 2.53(s, 3H), 2.14(s, 3H), 1.98(dd, 1H,J=5.3, 13.4 Hz)

MS (m/z): 453 (M⁺ +1).

EXAMPLE 38

Compound 38 (85 g, 64%) was obtained as pale yellow powder from Compoundl of Reference Example 9 and morpholine in a similar manner as inExample 37.

Melting point: 220°-223° C. (acetone-Et₂ O).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.33(d, 1H, J=8 Hz), 8.1-7.8(m, 3H),7.7-7.2(m, 4H), 6.81(dd, 1H, J=5, 7 Hz), 5.00(br.s, 2H), 3.85-3.6(m,4H), 3.57 (dd, 1H, J=7, 14 Hz), 3.1-2.5(m, 6H), ca. 2.17 (dd, 1H),2.15(s, 3H).

MS (m/z): 509 (M⁺ +1).

EXAMPLE 39

Compound 39 (66 mg, 63%) was obtained from 118 mg (0.2 mmol) of Compoundl of Reference Example 9 and 0.10 ml (1.0 mmol) of thiomorpholine in asimilar manner as in Example 37 as pale yellow powder showing a meltingpoint of 222° to 230° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.00-2.28(m, 1H), 2.12(s, 3H), 2.60-3.60(m,11H), 4.98(s, 2H), 5.26(s, 1H), 6.85(dd, 1H, J=5, 7 Hz), 7.16-7.84(m,5H), 7.92-8.16(m, 2H), 8.33(s, 1H), 9.32(d, 1H, J=8 Hz).

MS (m/e): 525 (M⁺ +1).

EXAMPLE 40

Compound 40 (127 mg, 100%) was obtained from 118 mg (0.2 mmol) ofCompound l of Reference Example 9 and 86 mg (1 mmol) of piperazine in asimilar manner as in Example 37 as yellowish green powder showing amelting point of 169° to 175° C.

NMR (DMSO-d₆) δ: 1.90-2.20(m, 1H), 2.12(s, 3H), 2.40-3.88(m, 11H),5.00(s, 2H), 5.40(s, 1H), 6.96-7.60(m, 4H), 7.81(d, 1H, J=7 Hz), 8.02(t,2H, J=7 Hz), 8.56(s, 1H), 9.20(d, 1H, J=8 Hz).

MS (m/e): 508 (M⁺ +1).

EXAMPLE 41

Compound 41 (37 mg, 37%) was obtained from 118 mg (0.2 mmol) of Compoundl of Reference Example 9 and 0.098 mg (1 mmol) of piperazine in asimilar manner as in Example 37 as brown powder showing a melting pointof 230° to 238° C.

NMR (DMSO-d₆) δ: 1.36-2.24(m, 7H), 2.16(s, 3H), 3.00-3.92(m, 7H),5.02(s, 2H), 6.28(br.s, 1H), 7.16-7.80(m, 6H), 7.92-8.12(m, 2H),8.63(br.s, 1H), 9.20(d, 1H, J=8 Hz).

MS (m/e): 507 (MH⁺).

EXAMPLE 42

Compound 42 (283 mg, 59.2%) was obtained from 593 mg (1 mmol) ofCompound l of Reference Example 9 and 0.75 mg (10 mmol) of allylamine ina similar manner as in Example 37 as brown powder showing a meltingpoint of 165° to 205° C.

NMR (CDCl₃) δ: 1.88-2.20(m, 1H), 2.04(s, 3H), 2.68-3.04(m, 1H), 2.92(d,1H, J=12 Hz), 3.22(d, 1H, J=12 Hz), 3.36-3.64(m, 2H), 4.36(d, 1H, J=17Hz) 4.56(d, 1H, J=17 Hz), 5.04(s, 1H), 5.12-5.24(m, 2H), 5.80-6.20(m,1H), 6.48(dd, 1H, J=5, 7 Hz), 6.92-7.60(m, 5H), 7.83(d, 1H, J=7 Hz),7.96(d, 1H, J=8 Hz), 8.70(d, 1H, J=8 Hz).

MS (m/e): 478 (M⁺).

EXAMPLE 43

In a solvent mixture of 2 ml of THF and 0.2 ml of pyridine was dissolved53.2 mg (0.1 mmol) of Compound 42, and 0.5 ml (0.1 mmol) of 0.05 g/mlTHF solution of osmium acid was added to the solution. After stirring atroom temperature for 30 minutes, a solution of 46 mg (0.44 mmol) ofsodium hydrogensulfite in 2 ml of water and 2 ml of pyridine was addedto the reaction mixture, followed by stirring at room temperature forone hour.

To the reaction solution was added 20 ml of THF, and the mixture waswashed with saturated NaCl aqueous solution and then dried overanhydrous magnesium sulfate. The solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(MeOH--CHCl₃ -28% NH₄ OH=5:95:0.5) and then dissolved in THF. 1.7NHCl/AcOEt was added to the solution to convert the product into thehydrochloride, whereby 22 mg (40%) of Compound 43 was obtained as brownpowder showing a melting point of 244° to 250° C.

NMR (DMSO-d₆ +D₂ O) δ: 2.16, 2.17(two s, 3H), 2.20 (dd, 1H, J=5, 14 Hz),3.09-3.15(m, 1H), 3.33-3.68 (m, 6H), 4.00-4.07(m, 1H), 4.98(d, 1H, J=18Hz), 5.06(d, 1H, J=18 Hz), 7.02-7.87(m, 1H), 7.32(t, 1H, J=8 Hz),7.40(t, 1H, J=7 Hz), 7.50-7.56(m, 2H), 7.68(d, 1H, J=8 Hz), 7.99(d, 1H,J=8 Hz), 8.08(d, 1H, J=8 Hz), 9.19(d, 1H, J=8 Hz).

MS (m/e): 513 (MH⁺).

EXAMPLE 44

Compound 44 (43 mg, 30%) was obtained as pale yellow powder fromCompound l of Reference Example 9 and guanidine hydrochloride in asimilar manner as in Example 37.

Melting point: 280°-285° C. (CH₃ OH--Et₂ O).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.33(d, 1H, J=8 Hz), 8.35-7.85(m, 3H),7.8-7.2(m, 4H), 7.10(dd, 1H, J=5, 7 Hz), 5.03(br. s, 2H), 3.80(br. s,2H), 3.12(dd, 1H, J=7, 14 Hz), 2.29(dd, 1H, J=5, 14 Hz), 2.22 (s, 3H).

MS (m/z): 463 (M⁺ -17).

EXAMPLE 45

To a solution of 237 mg (0.4 mmol) of Compound l of Reference Example 9in 5 ml of anhydrous THF was added 1.5 ml of 15% sodium methylatesolution, followed by stirring at room temperature for 7 hours. To thereaction mixture was added 30 ml of THF, and the resulting solution waswashed with an acid and an alkali. The solvent was removed under reducedpressure, and the residue was purified by silica gel columnchromatography (chloroform-methanol) to give 96 mg (51%) of Compound 45as pale yellow powder.

Melting point: 201°-205° C.

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.30(d, 1H, J=8 Hz), 8.15-7.8(m, 2H),7.7-7.2(m, 5H), 6.87(dd, 1H, J=5, 7 Hz), 5.08(d, 1H, J=13 Hz), 4.89(d,1H, J=13 Hz), 3.41(dd, 1H, J=7, 14 Hz), 3.30(d, 1H, J=13 Hz), 3.03(d,1H, J=13 Hz), 2.36(s, 3H), 2.23(s, 3H).

MS (m/z): 470 (M⁺ -1).

EXAMPLE 46

A solution of 188 mg (0.4 mmol) of Compound 45 in 5 ml of distilled THFwas ice cooled, and 2 ml of an aqueous solution of 90 mg (0.42 mmol) ofsodium periodate was added thereto, followed by stirring for 7 hoursunder ice cooling. To the reaction mixture was added 30 ml of THF, andthe resulting solution was washed with saturated sodium chloride aqueoussolution and then dried over anhydrous sodium sulfate. The solvent wasremoved under reduced pressure, and the residue was purified by silicagel column chromatography (chloroform-methanol) to give 163 mg (84%) ofCompound 46 as pale yellow powder.

Melting point: 235°-240° C.

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.31(d, 1H, J=8 Hz), 8.2-7.85(m, 2H),7.8-7.2(m, 5H), 7.2-6.9(m, 1H), 5.02(br. s, 2H), 3.9-3.2(m, 3H),2.89+2.87(2s, 3H), ca. 2.4(m, 1H), 2.27+2.19(2s, 3H).

MS (m/z): 486 (M⁺ +1).

EXAMPLE 47

To a suspension of 3.96 g (9 mmol) of Compound 20 in 200 ml of methanolwere added 2.05 g (9 mmol) of periodic acid and then 50 ml of THF,followed by stirring at room temperature for 2.5 hours. The solvent inthe reaction mixture was removed under reduced pressure and the residuewas dissolved in THF. The THF solution was washed with an acid and analkali. The solvent was removed under reduced pressure, and the residuewas purified by silica gel column chromatography (chloroform-methanol)to give 3.03 g (83%) of Compound 47 as pale yellow powder.

Melting point: 231°-234° C. (CH₂ Cl₂).

¹ H-NMR (DMSO-d₆) δ: 9.30(d, 1H, J=7.9 Hz), 8.1-7.8 (m, 3H), 7.6-7.3(m,5H), 4.98(s, 2H), 3.72(dd, 1H, J=7.0, 19.3 Hz), 2.77(d, 1H, J=3 Hz),2.48(s, 3H).

MS (m/z): 407 (M⁺).

IR (KBr): 3420, 1770, 1680, 1635, 1595, 1460, 740 cm⁻¹.

EXAMPLE 48

To a solution of 340 mg (0.83 mmol) of Compound 47 in 5 ml of THF and 5ml of EtOH was added 158 mg (4.17 mmol) of NaBH₄, followed by stirringat room temperature for one hour. To the reaction mixture was added 50ml of THF, and the resulting solution was washed with an acid and analkali and then dried over Na₂ SO₄. The solvent was removed underreduced pressure, and the solid residue obtained was recrystallized frommethanol to give 276 mg (81%) of pale yellow Compound 48.

Melting point: 251°-254° C.

¹ H-NMR (DMSO-d₆) δ: 9.22(d, 1H, J=7.8 Hz), 8.1-7.75 (m, 3H),7.55-7.25(m, 4H), 7.03(dd, 1H, J=4.5, 7.3 Hz), 5.77(d, 1H, J=6.2 Hz,exchangeable), 5.03(d, 1H, J=17.7 Hz), 4.98(d, 1H, J=17.7 Hz), 4.56(m,1H), 3.14(m, 1H), 2.29(s, 3H), 1.75(m, 1H).

MS (m/z): 409 (M⁺).

EXAMPLE 49

In 400 ml of THF was dissolved 4.67 g (10 mmol) of K-252, and 50 ml ofTHF solution of 0.38 g (10 mmol) of lithium aluminum hydride was addedto the solution at -20° C., followed by stirring at the same temperaturefor one hour. After 3N hydrochloric acid aqueous solution was added toadjust the pH to 2, the reaction mixture was filtered through Celite.The filtrate was washed with saturated sodium chloride aqueous solutionand then dried over anhydrous magnesium sulfate. The solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (chloroform) to give 1.56 g (35.7%) ofCompound 49 as pale yellow powder showing a melting point higher than300° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.04-2.48(m, 1H), 2.24(s, 3H), 3.08-3.76(1H),4.90(br. s, 2H), 6.91(dd, 1H, J=5, 7 Hz), 7.08-7.60(m, 5H), 7.76-8.08(m,2H), 9.19(d, 1H, J=8 Hz), 10.10(s, 1H).

MS (m/z): 437 (M⁺).

EXAMPLE 50

In 5 ml of THF and 0.5 ml of water was dissolved 100 mg (0.23 mmol) ofCompound 49, and 79 mg (1.1 mmol) of hydroxylamine hydrochloride wasadded to the solution, followed by stirring for one day. After thesolvent was distilled off under reduced pressure, the residue waspurified by silica gel column chromatography (1% methanol-chloroform) togive 85 mg (82%) of Compound 50 as pale yellow powder showing a meltingpoint of 245° to 256° C.

NMR (DMSO-d₆) δ: 1.98-2.30(m, 1H), 2.20(s, 3H), 3.16-3.70(m, 1H),5.03(s, 2H), 6.84-7.08(m, 1H), 7.16-8.20(m, 8H), 8.58(s, 1H), 9.26(d,1H, J=8 Hz).

MS (m/e): 452 (M⁺).

EXAMPLE 51

Compound 51 (55 mg, 53%) was obtained from 87 mg (0.2 mmol) of Compound49 and 181 mg (1.0 mmol) of 2-hydrazino-2-imidazoline hydrobromide in asimilar manner as in Example 50 as pale yellow powder showing a meltingpoint higher than 300° C.

NMR (DMSO-d₆) δ: 1.68-2.30(m, 1H), 2.08(s, 3H), 3.00-3.70(1H), 5.00(s,2H), 5.96(s, 1H), 7.00-8.12(m, 8H), 8.56(s, 1H), 9.21(d, 1H, J=8 Hz).

MS (m/e): 520 (M⁺ +1).

EXAMPLE 52

Compound 52 (60 mg, 60%) was obtained from 87 mg (0.2 mmol) of Compound49 and 264 mg (1.0 mmol) of aminoguanidine sulfate in a similar manneras in Example 50 as pale yellow powder showing a melting point higherthan 300° C.

NMR (DMSO-d₆) δ: 1.96-2.30(m, 1H), 2.15(s, 3H), 3.04-3.64(m, 1H),5.02(br. s, 1H), 6.44(s, 1H), 7.00-8.20(m, 8H), 8.60(s, 1H), 9.22(d, 1H,J=8 Hz).

MS (m/e): 494 (M⁺ +1).

EXAMPLE 53

To a solution of 132 mg (0.3 mmol) of Compound 20 in anhydrous THF wereadded 146 mg (0.9 mmol) of 1,1'-carbonyldiimidazole and 42 μl (0.3 mmol)of triethylamine, followed by stirring at room temperature overnight. Tothe reaction mixture was added 50 ml of THF, and the resulting solutionwas washed with an acid and an alkali. The solvent was removed underreduced pressure, and the solid residue was recrystallized fromTHF-dichloromethane to give 75 mg (54%) of Compound 53 as whitecrystals.

Melting point: 257°-260° C.

¹ H-NMR (DMSO-d₆) δ: 9.23(d, 1H, J=7.9 Hz), 8.15-7.75 (m, 3H),7.55-7.25(m, 4H), 7.21(dd, 1H, J=4.3, 7.1 Hz), 5.21(d, 1H, J=9.4 Hz),5.02(s, 2H), 4.83 (d, 1H, J=9.4 Hz), 3.40(dd, 1H, J=7.1, 14.7 Hz),2.53(dd, 1H, J=4.3, 14.7 Hz), 2.46(s, 3H).

MS (m/z): 466 (M⁺ +1).

IR (KBr): 3450, 1810, 1680, 1640, 1590, 1460, 745 cm⁻¹.

EXAMPLE 54

Compound 54 (75 mg, 52%) was obtained as pale yellow powder fromCompound 20 and 1,1'-thiocarbonyldiimidazole in a similar manner as inExample 53.

Melting point: 255°-258° C. (THF-CH₃ OH).

¹ H-NMR (DMSO-d₆) δ: 9.24(d, 1H, J=7.9 Hz), 8.15-7.75 (m, 3H),7.6-7.3(m, 4H), 7.23(dd, 1H, J=4.2, 7.1 Hz), 5.52(d, 1H, J=10.0 Hz),5.09(d, 1H, J=10.0 Hz), 5.02(s, 2H), 3.48(dd, 1H, J=7.1, 14.8 Hz),2.65(dd, 1H, J=4.2, 14.8 Hz), 2.44(s, 3H).

MS (m/z): 481 (M⁺).

IR (KBr): 3440, 1680, 1640, 1590, 1460, 1315, 745 cm⁻¹.

EXAMPLE 55

To a solution of 132 mg (0.3 mmol) of Compound 20 in 2 ml of pyridinewas added 33 μl (0.45 mmol) of thionyl chloride, followed by stirring atroom temperature for one hour. To the reaction mixture was added 50 mlof THF, and the resulting solution was washed with 5% citric acidaqueous solution and saturated sodium chloride aqueous solution and thendried over anhydrous sodium sulfate. The solvent was removed underreduced pressure, and the solid residue was recrystallized from methanolto give 97 mg (71%) of Compound 55 as pale yellow powder.

Melting point: 256°-259° C.

¹ H-NMR (DMSO-d₆) δ: 9.3-9.05(m, 1H), 8.3-7.7(m, 3H), 7.7-7.2(m, 5H),5.75-4.85(m, 4H), 3.5-3.25(m, 1H), 2.65-2.35(m, 1H), 2.40+2.38(2s, 3H).

MS (m/z): 486 (M⁺ +1).

EXAMPLE 56

In 7 ml of sulfolane was dissolved 131 mg (0.3 mmol) of the free amineof Compound 30, and 0.057 ml (0.45 mmol) of trimethyl orthoacetate and 3mg of camphor sulfonic acid were added to the solution, followed byheating at 90° C. for 2.5 hours. To the reaction solution was added 20ml of water. The precipitates were separated by filtration and purifiedby silica gel chromatography (1% MeOH--CHCl₃) to give 45 mg (32%) ofCompound 56 as brown powder showing a melting point of 227° to 233° C.

NMR (CDCl₃) δ: 1.84(s, 3H), 2.44(s, 3H), 2.72(dd, 1H, J=5, 14 Hz),3.20(dd, 1H, J=7, 14 Hz), 4.01 (d, 1H, J=13 Hz), 4.64(d, 1H, J=13 Hz),5.16(s, 2H), 6.56(br. s, 1H), 6.94(dd, 1H, J=5, 7 Hz), 7.44-7.84(m, 6H),8.00-8.16(m, 1H), 9.46(d, 1H, J=8 Hz).

MS (m/z): 462 (M⁺).

EXAMPLE 57

Compound 57 (29 mg, 21%) was obtained as pale yellow powder from thefree amine of Compound 30 and 1,1'-carbonyldiimidazole in a similarmanner as in Example 53.

Melting point: >300° C. (CH₂ Cl₂ --CH₃ OH).

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.36(d, 1H, J=8 Hz), 8.2-7.05(m, 8H),5.06(br. s, 2H), 4.18(d, 1H, J=10 Hz), 3.86(d, 1H, J=10 Hz), 3.20(dd,1H), 2.41 (s, 3H).

MS (m/z): 465 (M⁺ +1).

IR (KBr): 3430, 1765, 1670, 1640, 1590, 1460, 745 cm⁻¹.

EXAMPLE 58

In 5 ml of DMF was dissolved 120 mg (0.27 mmol) of the free amine ofCompound 30, and 98 mg (0.55 mmol) of thiocarbonyldiimidazole was addedto the solution under ice cooling, followed by stirring at the sametemperature for one hour. To the reaction solution was added 30 ml ofTHF, and the resulting solution was washed with saturated sodiumchloride aqueous solution and then dried over anhydrous magnesiumsulfate. The solvent was distilled off under reduced pressure to give120 mg (93%) of Compound 58 as pale yellow powder showing a meltingpoint higher than 300° C.

NMR (DMSO-d₆) δ: 2.10-2.64(m, 1H), 2.32(s, 3H), 3.00-3.52(m, 1H),4.05(d, 1H, J=11 Hz), 4.38(d, 1H, J=11 Hz), 5.02(s, 2H), 6.96-8.16(m,7H), 8.60(s, 1H), 9.21(d, 1H, J=8 Hz).

MS (m/e): 481 (M⁺ +1).

EXAMPLE 59

Compound 59 (245 mg, 58%) was obtained from 400 mg (0.88 mmol) ofCompound 37 and 287 mg (1.77 mmol) of 1,1'-carbonyldiimidazole in asimilar manner as in Example 53 as yellow powder showing a melting pointof 267° to 276° C.

NMR (DMSO-d₆) δ: 2.24-2.52(m, 1H), 2.34(s, 3H), 2.88 (s, 3H),3.08-3.40(m, 1H), 3.80(d, 1H, J=10 Hz), 4.16(d, 1H, J=10 Hz), 5.00(s,2H), 7.08-7.56(m, 5H), 7.64-7.84(m, 2H), 8.04(d, 1H, J=8 Hz), 8.56 (s,1H), 9.18(d, 1H, J=8 Hz).

MS (m/e): 479 (M⁺ +1).

EXAMPLE 60

Compound 60 (2.03 mg, 97%) was obtained from 2.04 mg (4.26 mmol) ofCompound 42 and 1.03 mg (6.39 mmol) of 1,1'-carbonyldiimidazole in asimilar manner as in Example 53 as pale green powder showing a meltingpoint of 230° to 237° C.

NMR (DMSO-d₆) δ: 2.24-2.60(m, 1H), 2.36(s, 3H), 3.16-3.24(m, 1H),3.82(d, 1H, J=10 Hz), 3.94(d, 2H, J=8 Hz), 4.14(d, 1H, J=10 Hz), 5.03(s,2H), 5.24-5.52(m, 2H), 5.76-6.20(m, 1H), 7.12-7.92 (m, 7H), 8.10(d, 1H,J=8 Hz), 8.64(s, 1H), 9.23 (d, 1H, J=8 Hz).

MS (m/e): 495 (M⁺ +1).

EXAMPLE 61

Compound 61 (95 mg, 59%) was obtained from 148 mg (0.3 mmol) of Compound60 in a similar manner as in Example 43 as yellowish brown powdershowing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.20-2.60(m, 1H), 2.40(s, 3H), 3.00-4.12(m, 7H),4.20-4.44(m, 1H), 5.02(s, 2H), 7.16-7.92(m, 7H), 8.09(br. d, 1H, J=8Hz), 8.62 (br. s, 1H), 9.23(d, 1H, J=8 Hz).

MS (m/e): 539 (M⁺ +1).

EXAMPLE 62

Compound 62 (42 mg, 69%) was obtained from 64 mg (0.12 mmol) of Compound61 in a similar manner as in Example 47 as pale yellow powder showing amelting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.10-2.60(m, 1H), 2.40(s, 3H), 3.04-3.60(m, 1H),3.68-4.60(m, 4H), 5.04(s, 2H), 7.08-8.24(m, 8H), 8.64(br. s, 1H),9.24(d, 1H, J=8 Hz), 9.64(s, 1H).

MS (m/e): 507 (M⁺ +1).

EXAMPLE 63

Compound 63 (12 mg, 30%) was obtained from 35 mg (0.07 mmol) of Compound62 and 93 mg (0.35 mmol) of aminoguanidine sulfate in a similar manneras in Example 50 as pale yellow powder showing a melting point higherthan 300° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.16-2.68(m, 1H), 2.40(s, 3H), 3.00-4.40(m, 5H),5.05(s, 2H), 7.04-8.20(m, 8H), 8.44(br. s, 1H), 9.34(d, 1H, J=8 Hz).

MS (m/e): 563 (M⁺ +1).

EXAMPLE 64

In 2 ml of DMF was dissolved 88 mg (0.18 mmol) of Compound 58, and 0.1ml of methyl iodide was added to the solution, followed by stirring atroom temperature for 2.5 hours. After the solvent was distilled offunder reduced pressure, the residue was purified by silica gel columnchromatography (CHCl₃) to give 14 mg (15.7%) of Compound 64 as yellowpowder showing a melting point of 223° to 225° C.

NMR (DMSO-d₆) δ: 2.08-2.44(m, 1H), 2.24(s, 3H), 2.30(s, 3H), 3.20(dd,1H, J=7, 14 Hz), 4.06(d, 1H, J=14 Hz), 4.57(d, 1H, J=14 Hz), 5.02(s,2H), 7.12-8.20(m, 7H), 8.63(s, 1H), 9.24(d, 1H, J=8 Hz).

MS (m/e): 494 (M⁺).

EXAMPLE 65

To 50 ml of pyridine was added 3 g (30 mmol) of chromic acid under icecooling, and after 10 minutes, 30 ml of a solution of 2.9 g (6.2 mmol)of K-252 in pyridine was added thereto, followed by stirring at roomtemperature for one day. The reaction mixture was filtered throughCelite, and THF was added to the filtrate. The mixture was washed withsaturated sodium chloride aqueous solution and then dried over anhydrousmagnesium sulfate (MgSO₄). The solvent was distilled off under reducedpressure, and the residue obtained was purified by silica gel columnchromatography (chloroform) and recrystallized from chloroform to give2.53 g (85%) of Compound 65 as yellow powder showing a melting point of288° to 290° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.18(s, 3H), 2.38(dd, 1H, J=5, 14 Hz), 3.37(dd,1H, J=6, 14 Hz), 4.00(s, 3H), 6.05(br. s, 1H), 6.95(dd, 1H, J=5, 6 Hz),7.20-8.04(m, 6H), 9.08(d, 1H, J=8 Hz), 9.28(d, 1H, J=8 Hz), 10.16(s,1H).

MS (m/e): 481 (M⁺).

EXAMPLE 66

Compound 66 (110 mg, 97%) was obtained from 150 mg (0.25 mmol) ofCompound o of Reference Example 12 in a similar manner as in Example 34as yellow needles showing a melting point of 274° to 280° C.

NMR (CDCl₃ +DMSO-d₆) δ: 1.72-2.20(m, 1H), 2.16(s, 3H), 2.90-4.40(m, 6H),6.84-7.00(m, 1H), 7.20-8.08(m, 7H), 9.09(d, 1H, J=8 Hz), 9.28(d, 1H, J=8Hz).

MS (m/e): 453 (M⁺ +1).

EXAMPLE 67

Compound 67 (1.14 mg, 84%) was obtained from 1.44 mg (3 mmol) ofCompound 65 in a similar manner as in Example 22 as yellow powdershowing a melting point of 278° to 280° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.13(dd, 1H, J=5, 14 Hz), 2.18(s, 3H),3.00-3.36(m, 1H), 3.76-4.04(m, 2H), 6.76-7.00(m, 1H), 7.20-7.84(m, 5H),8.01(d, 1H, J=8 Hz), 9.12(d, 1H, J=8 Hz), 9.30(d, 1H, J=8 Hz).

MS (m/e): 453 (M⁺).

EXAMPLE 68

In 10 ml of DMF was dissolved 962 mg (2 mmol) of Compound 65, and 80 mg(2 mmol) of 60% sodium hydride was added to the solution under icecooling. After 10 minutes, 0.17 ml (2 mmol) of 1,1-dibromomethane wasadded thereto, followed by stirring at room temperature for 2 days. Tothe reaction solution was added 2 ml of saturated NH₄ Cl aqueoussolution, and the mixture was extracted with THF. After washing withsaturated NaCl aqueous solution, the extract was dried over anhydrousmagnesium sulfate. The solvent was distilled off under reduced pressure,and the residue was purified by silica gel column chromatography (CHCl₃)to give 503 mg (43%) of Compound 68 as yellow prisms showing a meltingpoint of 248° to 249° C.

NMR (CDCl₃) δ: 2.20(s, 3H), 2.39(dd, 1H, J=5, 14 Hz), 3.35(dd, 1H, J=7,14 Hz), 3.68 (t, 2H, J=7 Hz), 3.79(s, 1H), 4.10(t, 2H, J=7 Hz), 6.88(dd,1H, J=5, 7 Hz), 7.24-7.90(m, 6H), 9.12(d, 1H, J=8 Hz), 9.33(d, 1H, J=8Hz).

MS (m/e): 588 (M⁺).

EXAMPLE 69

In DMF was dissolved 480 mg (0.81 mmol) of Compound 68, and 1.32 g (16.2mmol) of dimethylamine hydrochloride and 24 ml of DBU were added to thesolution, followed by stirring at room temperature for one day. Thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (MeOH--CHCl₃ -28% NH₄OH=2.5:97.5:0.1) and then converted into the hydrochloride with 1.7NHCl/AcOEt to give 459 mg (78%) of Compound 69 as yellow powder showing amelting point of 248° to 249° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.12-2.40(m, 1H), 2.22(s, 3H), 3.02(s, 6H),3.32-3.84(m, 3H), 4.01(s, 3H), 4.08-4.32(m, 2H), 6.44(br. s, 1H),7.00-7.28 (m, 1H), 7.32-8.16(m, 6H), 9.08(d, 1H, J=8 Hz), 9.28(d, 1H,J=8 Hz).

MS (m/e): 553 (MH⁺).

EXAMPLE 70

Compound 70 (80 mg, 30.4%) was obtained from 260 mg (0.47 mmol) of thefree amine of Compound 69 ml in a similar manner as in Example 22 asyellow powder showing a melting point of 221° to 230° C.

NMR (DMSO-d₆) δ: 1.84-2.20(m, 1H), 2.19(s, 3H), 2.52(s, 6H),3.00-3.60(m, 3H), 3.72-3.96(m, 2H), 4.00-4.24(m, 2H), 5.18(br. s, 1H),5.56(s, 1H), 6.94-7.16(m, 1H), 7.24-8.12(m, 6H), 9.00(d, 1H, J=8 Hz),9.20(d, 1H, J=8 Hz).

MS (m/e): 525 (MH⁺).

EXAMPLE 71

In 2 ml of dioxane was dissolved 45 mg (0.1 mmol) of Compound 66, and0.25 ml (5 mmol) of hydrazine hydrate was added to the solution,followed by heating at 100° C. for 2 hours. The solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (MeOH--CHCl₃ -28% NH₄ OH=5:95:0.5) and thenconverted into the hydrochloride with 1.7N HCl/AcOET to give 43 mg(85.5%) of Compound 71 as yellow prisms showing a melting point of 270°to 275° C.

NMR (DMSO-d₆ +D₂ O) δ: 2.12-2.44(m, 1H), 2.24(s, 3H), 3.20-3.72(m, 3H),7.16-8.24(m, 7H), 9.10(d, 1H, J=8 Hz), 9.31(d, 1H, J=8 Hz).

MS (m/e): 468 (MH⁺).

EXAMPLE 72

Compound 72 (472 mg, 62%) was obtained from 743 mg (1.64 mmol) ofCompound 67 and 3.97 mg of hydrazine hydrate in a similar manner as inExample 71 as yellow powder showing a melting point of 245° to 260° C.

NMR (DMSO-d₆) δ: 2.19(dd, 1H, J=5, 14 Hz), 2.16(s, 3H), 3.00-3.40(m,3H), 3.80(br. s, 2H), 4.96 (br. s, 1H), 5.44(br. s, 1H), 6.88-7.12 (m,1H), 7.24-8.20(m, 6H), 9.04(d, 1H, J=8 Hz), 9.04(d, 1H, J=8 Hz).

MS (m/e): 469 (M⁺ +1).

EXAMPLE 73

In 5 ml of DMF was dissolved 285 mg (0.63 mmol) of Compound 67, and 0.42ml of ethylenediamine and 1.91 ml of DBU were added to the solution,followed by heating at 70° C. for 4 hours. The solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (CHCl₃ --MeOH-28% NH₄ OH=100:5:0.1) and then convertedinto the hydrochloride with 1.7N HCl/AcOEt to give 335 mg (99%) ofCompound 73 as yellow powder showing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.01(dd, 1H, J=5, 14 Hz), 2.20(s, 3H), 3.00-3.50(m,4H), 3.68-4.20(m, 4H), 5.60 (br. s, 1H), 6.96-7.20(m, 1H), 7.24-7.76(m,4H), 7.92(d, 1H, J=8 Hz), 8.06(d, 1H, J=8 Hz), 8.20 (br. s, 2H), 9.04(d,1H, J=8 Hz), 9.20 (d, 1H, J=8 Hz).

MS (m/e): 496 (M⁺).

EXAMPLE 74

Compound 74 (65 mg, 68%) was obtained from 93 mg (0.2 mmol) of Compound57 in a similar manner as in Example 65 as yellow powder showing amelting point higher than 300° C.

NMR (DMSO-d₆) δ: 2.33-2.54(m, 1H), 2.38(s, 3H), 3.27-3.41(m, 1H),3.81(d, 1H, J=10 Hz), 4.14 (d, 1H, J=10 Hz), 7.24(dd, 1H, J=5, 7 Hz),7.42 (t, 1H, J=8 Hz), 7.56(t, 1H, J=8 Hz), 7.62(t, 1H, J=8 Hz), 7.84(d,2H, J=8 Hz), 8.04(s, 1H), 9.02 (d, 1H, J=8 Hz), 9.22(d, 1H, J=8 Hz),11.16(s, 1H).

MS (m/e): 479 (M⁺).

EXAMPLE 75

Compound 75 (544 mg, 66.3%) was obtained from 800 mg (1.67 mmol) ofCompound 59 in a similar manner as in Example 65 as yellowish brownpowder showing a melting point of 290° to 292° C.

NMR (DMSO-d₆) δ: 2.24-2.60(m, 1H), 2.34(s, 3H), 2.86(s, 3H),3.12-3.48(m, 1H), 3.82(d, 1H, J=12 Hz), 3.18(d, 1H, J=12 Hz),7.16-7.96(m, 1H), 9.00(d, 1H, J=8 Hz), 9.18(d, 1H, J=8 Hz), 10.64 (s,1H).

MS (m/e): 493 (M⁺ +1).

EXAMPLE 76

In 30 ml of pyridine was dissolved 2.4 g (5 mmol) of Compound 1, and1.53 ml of acetic anhydride was added to the solution. The mixture wasstirred at room temperature for 3 hours. The solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (2% MeOH--CHCl₃) to give 630 mg (23%) of thediacetyl compound. The compound obtained was oxidized in a similarmanner as in Example 65 to give 522 mg (81%) of the imide compound. In 1ml of DMF was dissolved 56.6 g (0.1 mmol) of the imide compound, and 0.2ml of 28% NH₄ OH aqueous solution was added to the solution, followed bystirring at room temperature for one day. The solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (2% MeOH--CHCl₃) to give 36 mg (75%) of Compound76 as brown powder showing a melting point higher than 300° C.

NMR (CDCl₃ +DMSO-d₆) δ: 2.16-2.44(m, 1H), 2.24(s, 3H), 3.24-3.52(m, 2H),6.34(br, s, 1H), 6.92-7.12(m, 1H), 7.24-8.20(m, 6H), 8.92-9.40(m, 3H),10.86(br. s, 1H).

MS (m/e): 483 (M⁺ +1).

EXAMPLE 77

Compound (I) was examined for C-kinase inhibitory activity and cellgrowth inhibitory activity by the following method. The results areshown in Table 4.

C-Kinase inhibitory activity test:

C-Kinase inhibitory activity of representative Compounds (I) wasmeasured in accordance with the method of Y. Nishizuka, et al. [J. Biol.Chem., 257, 13341 (1982)]. The test was carried out on test compounds atvaried concentrations, and the concentration at which the enzymeactivity was inhibited 50% (IC₅₀) was determined.

(1) MCF 7 cell growth inhibition test:

MCF 7 cells (4.5×10⁴ cells/ml) prepared in RPMI 1640 medium containing10% fetal calf serum, 10 μg/ml insulin and 10⁻⁸ M estradiol are put intowells of a 96-well microtiter plate in the amount of 0.1 ml per eachwell. After incubation at 37° C. overnight in CO₂ -incubator, 0.05 mleach of a test sample appropriately diluted with the culture medium isadded to each well. In the case of contact for 72 hours, the cells arefurther cultured in CO₂ -incubator, and then the culture supernatant isremoved and the wells are washed once with PBS(-). Thereafter, 0.1 ml offresh medium is added to each well, followed by culturing at 37° C. for72 hours in CO₂ -incubator. After the culture supernatant is removed,0.1 ml of the culture medium containing 0.02% Neutral Red is added toeach well, followed by culturing at 37° C. for one hour in CO₂-incubator, whereby the cells are stained. Then, the culture supernatantis removed and the cells are washed once with physiological saline. Thepigment is extracted with 0.001N HCl/30% ethanol and absorption ismeasured at 550 nm with a microplate reader. By comparing the absorptionof intact cells with those of the cells treated with a test compound inknown concentrations, the concentration of the test compound at whichgrowth of the cells is inhibited 50% is calculated as IC₅₀.

(2) PC-10 cell growth inhibition test:

PC-10 cells (5×10⁴ cells/ml) prepared in MEM medium containing 10% fetalcalf serum and 2 mM glutamine are put into wells of a 96-well microtiterplate in the amount of 0.1 ml per each well.

Thereafter, the test is carried out in the same manner as in the MCF 7cell growth inhibition test.

(3) HeLaS₃ cell growth inhibition test:

HeLaS₃ cells (3×10⁴ cells/ml) prepared in MEM medium containing 10%fetal calf serum and 2 mM glutamine are put into wells of a 96-wellmicrotiter plate in the amount of 0.1 ml per each well.

Thereafter, the test is carried out in the same manner as in (1).

(4) COLO320DM cell growth inhibition test:

COLO320DM cells (10⁵ cells/ml) prepared in RPMI 1640 medium containing10% fetal calf serum, 100 U/ml penicillin and 100 μg/ml streptomycin areput into wells of a 96-well microtiter plate in the amount of 0.1 ml pereach well. Thereafter, the test is carried out in the same manner as in(1) except that the cells are counted with a microcell counter. Bycomparing the number of intact cells with those of the cells treatedwith a test compound in known concentrations, the concentration of thetest compound at which growth of the cells is inhibited 50% iscalculated as IC₅₀.

                  TABLE 4                                                         ______________________________________                                        C-Kinase Inhibitory Activity and                                              Cell Growth Inhibitory Activity of the Compounds                              Com-                                                                          pound IC.sub.50 (μg/ml)                                                    No.   C-Kinase HeLaS.sub.3                                                                            PC-10 MCF-7 COLO320DM                                 ______________________________________                                         1    0.005    0.59     0.14                                                   2    0.003    0.22     0.03                                                   3    0.018    0.95     0.68                                                   4    0.13     0.6      >1                                                     5    0.034    0.51     0.84                                                   7    0.028    0.15     0.76                                                   9    0.175    0.01           0.13  0.05                                      10    0.02     0.07           0.95  0.10                                      11    --       0.10           0.47  0.16                                      12    --       0.17           0.36  0.11                                      15    0.009    0.48     0.11                                                  16    0.005    0.44           0.84                                            20    0.02     0.07     0.11  0.55  0.05                                      21    0.01     0.46           0.76  0.29                                      23    0.038    0.15           0.34  0.22                                      24    --       0.06           0.19  0.06                                      26    --       0.03           0.26  0.03                                      27    0.017    0.53           >1    >1                                        28    --       0.20           1.0   0.32                                      29    0.06     0.44     0.71                                                  30    0.015    0.07     0.04                                                  31    --       0.08           0.8   0.31                                      32    0.01     0.22           0.74  >1                                        33    --       0.08           0.26  1.0                                       34    --       0.03           0.24  0.17                                      35    --       0.05           0.27  0.26                                      36    0.02     0.08           0.52  0.12                                      38    0.10     0.18                 0.30                                      39    --       0.14           >1    0.38                                      40    --       0.38           >1    >1                                        41    --       0.06           >1    0.07                                      43    0.02     0.19           0.52  >1                                        45    0.11     0.55     >1                                                    46    0.06     >1       0.27                                                  48    0.022    0.72     0.48                                                  50    0.013    0.22                 0.17                                      51    0.031    0.23           0.50  1.0                                       52    0.021    0.46           0.40  >1                                        54    0.015    0.28     0.11  1.0   0.08                                      55    0.038    0.14     0.64                                                  56    0.006    0.07           0.40  1.0                                       57    0.02     0.04     0.10  0.15  0.37                                      58    0.017    0.28     0.69                                                  59    0.046    0.17           1.0   0.39                                      63    0.053    0.20           0.45  0.64                                      67    0.016    0.95                 0.18                                      69    --       0.88           >1    0.68                                      70    --       0.50           0.45  0.41                                      71    --       >1             1.0   >1                                        72    0.22     0.24           0.2   0.37                                      75    --       0.27                 0.18                                      76    --       0.40           0.62  0.06                                      ______________________________________                                    

REFERENCE EXAMPLE 1

To a solution of 4.53 g (10 mmol) of Compound KT-5556 in 50 ml ofanhydrous pyridine was added 1.42 ml (15 mmol) of acetic anhydride,followed by stirring at room temperature for one hour. The solvent inthe reaction mixture was distilled off under reduced pressure and 50 mlof 1N hydrochloric acid was added to the residue, followed by stirring.Insoluble matters were separated by filtration, washed with 1Nhydrochloric acid and then with water, and dried under reduced pressureto give 4.79 g (97%) of Compound a as pale yellow powder.

Melting point: 267°-270° C.

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.36(d, 1H, J=8 Hz), 8.2-7.7(m, 3H),7.7-7.25(m, 4H), 7.25(dd, 1H, J=5, 7Hz), 5.07(s, 2H), 3.98(dd, 1H, J=7,14 Hz), 2.35(s, 3H), 2.12(dd, 1H, J=5, 14 Hz), 1.72(s, 3H).

IR (KBr): 3430, 1750, 1680, 1640, 1590, 1460, 1235, 745 cm⁻¹.

REFERENCE EXAMPLE 2

In 10 ml of THF was dissolved 2 g (4.2 mmol) of K-252, and 4 ml ofacetic anhydride and 2.6 g of dimethylaminopyridine were added to thesolution, followed by stirring at room temperature overnight. Thereaction solution was washed successively with 2% hydrochloric acidaqueous solution and saturated sodium chloride aqueous solution and thendried over anhydrous magnesium sulfate. The solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (CHCl₃) to give 2.12 g (94%) of Compound b as paleyellow powder.

NMR (CDCl₃) δ: 1.76(s, 3H), 2.03(dd, 1H, J=5, 14 Hz), 2.16(s, 3H),2.56(s, 3H), 3.86(dd, 1H, J=7, 14 Hz), 3.98(s, 3H), 5.07(s, 2H),6.93(dd, 1H, J=5, 7 Hz), 7.14-7.66(m, 5H), 7.80-8.00(m, 2H), 9.02(d, 1H,J=8 Hz).

REFERENCE EXAMPLE 3

In 100 ml of sulfolane and 50 ml of chloroform was dissolved 5.51 g (10mmol) of Compound b, and 2.8 g (10.5 mmol) of nitroniumtetrafluoroborate was added to the solution. The mixture was heated at80° C. for 2 hours. After chloroform was distilled off under reducedpressure, 200 ml of water was added to the residue. The precipitateswere separated by suction filtration and washed with water and methanolto give a mixture of Compound V-1 (R¹⁵ =Me), Compound V-2 (R¹⁵ =Me) andCompound V-3 (R¹⁵ =Me).

The mixture obtained was then dissolved in 250 ml of DMF and 2 g of 10%palladium/carbon was added to the solution, followed by stirring at roomtemperature in a stream of hydrogen. After 2 hours, the reactionsolution was filtered through Celite and the solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: chloroform) and recrystallized from achloroform-ether solvent mixture to give 1.74 g (30%) of Compound c asyellow needles showing a melting point higher than 300° C. and 0.59 g(10%) of Compound d as yellow powder showing a melting point higher than300° C.

Further, 0.35 g (6.1%) of Compound e was obtained as yellow powdershowing a melting point higher than 300° C.

Compound c:

NMR (CDCl₃) δ: 1.79(s, 3H), 2.12(dd, 1H, J=5, 14 Hz), 2.28(s, 3H),2.83(s, 3H), 3.98(dd, 1H, J=7, 14 Hz), 4.03(s, 3H), 5.36(s, 2H),6.83-7.10(m, 2H), 7.23-7.66(m, 3H), 7.93(dd, 1H, J=2, 8 Hz), 8.60(dd,1H, J=2, 8 Hz), 8.54(d, 1H, J=2 Hz).

MS (m/e): 567 (M⁺ +1).

Compound d:

NMR (CDCl₃) δ: 1.74(s, 3H), 2.08(dd, 1H, J=5, 8 Hz), 2.15(s, 3H),2.71(s, 3H), 3.83(dd, 1H, J=7, 14Hz), 3.93(s, 3H), 5.00(br. s, 4H),5.32(s, 2H), 6.80-7.20(m, 3H), 7.28(br. s, 1H), 7.67 (d, 1H, J=8 Hz),7.70(d, 1H, J=8 Hz), 8.33(d, 1H, J=2 Hz).

MS (m/e): 582 (M⁺ +1).

Compound e:

NMR(DMSO-d₆) δ: 1.74(s, 3H), 2.08-2.40(m, 1H), 2.20(s, 3H), 2.60(s, 3H),3.80-4.12(m, 1H), 3.96(s, 3H), 5.04(br. s, 2H), 5.32(s, 2H),6.96-7.20(m, 2H), 7.32-7.88(m, 3H), 8.06(d, 1H, J=8 Hz), 8.72(dd, 1H,J=2, 8 Hz).

MS (m/e): 567 (M⁺ +1).

REFERENCE EXAMPLE 4

In 10 ml of dichloromethane was dissolved 110 mg (0.2 mmol) of Compoundb, and 133 mg (1 mmol) of aluminum chloride and 0.015 ml (0.2 mmol) ofacetyl chloride were added to the solution under ice cooling. Themixture was stirred at the same temperature for 2 hours, and 10 ml ofwater was added thereto, followed by extraction of the organic layer.The organic layer was washed with saturated sodium chloride aqueoussolution and then dried over anhydrous magnesium sulfate. The residuewas purified by silica gel column chromatography (chloroform) andrecrystallized from chloroform-methanol to give 60 mg (50.8%) ofCompound f as colorless prisms showing a melting point higher than 300°C. Further, 5 mg (4%) of Compound g was obtained as yellow prismsshowing a melting point higher than 300° C.

Compound f:

NMR (CDCl₃) δ: 1.76(s, 3H), 1.09(dd, 1H, J=5, 14 Hz), 2.28(s, 3H),2.52(s, 3H), 2.69(s, 3H), 3.93(dd, 1H, J=7, 14 Hz), 4.01(s, 3H), 5.20(s,3H), 6.89 (dd, 1H, J=5, 7 Hz), 7.28-7.72(m, 3H), 7.88-8.24 (m, 3H),9.68(s, 1H).

MS (m/e): 594 (M⁺ +1).

Compound g:

NMR (CDCl₃) δ: 1.82(s, 3H), 2.21(dd, 1H, J=5, 14 Hz), 2.34(s, 3H),2.75(s, 3H), 2.80(s, 3H), 2.82(s, 3H), 4.06(dd, 1H, J=7, 14 Hz), 4.07(s,3H), 5.40 (s, 2H), 7.03(dd, 1H, J=5, 7 Hz), 7.56(d, 1H, J=8 Hz), 8.01(d,1H, J=8 Hz), 8.24(d, 1H, J=8 Hz), 8.25(d, 1H, J=8 Hz), 8.60(s, 1H),9.84(d, 1H, J=2 Hz).

Ms (m/e): 636 (M⁺ +1).

REFERENCE EXAMPLE 5

In 30 ml of dichloromethane was dissolved 330 mg (0.6 mmol) of Compoundb, and 0.46 ml (4.2 mmol) of titanium tetrachloride and 0.11 ml (1.2mmol) of dichloromethane methyl ether were added to the solution underice cooling. The mixture was stirred at room temperature for 3 hours,and 10 ml of water was added thereto, followed by extraction. Theorganic layer was washed with saturated sodium chloride aqueous solutionand then dried over anhydrous magnesium sulfate. After the solvent wasdistilled off under reduced pressure, the residue was purified by silicagel column chromatography (chloroform) and recrystallized fromchloroform-methanol to give 130 mg (37%) of Compound h as colorlessprisms showing a melting point higher than 300° C.

Compound h:

NMR (DMSO-d₆) δ: 1.72(s, 3H), 2.04-2.36(m, 1H), 2.25(s, 3H), 2.68(s,3H), 3.80-4.08(m, 1H), 4.00(s, 3H), 5.43(s, 2H), 7.20-8.40(m, 7H),9.60(s, 1H), 10.16(s, 1H).

MS (m/e): 580 (M⁺ +1).

REFERENCE EXAMPLE 6

In a solvent mixture of 20 ml of methanol and 100 ml of chloroform wasdissolved 2.51 g (4.3 mmol) of Compound h, and 488 mg (12.4 mmol) ofsodium borohydride was added to the solution under ice cooling. Themixture was stirred at the same temperature for 30 minutes. 3NHydrochloric acid aqueous solution was added to adjust the pH to 2,followed by extraction. The organic layer was washed successively withsaturated sodium bicarbonate aqueous solution and saturated sodiumchloride aqueous solution and then dried over anhydrous magnesiumsulfate. The residue was triturated with ether to give 1.8 g (72%) ofCompound i as pale yellow powder showing a melting point of 270° to 277°C.

NMR (CDCl₃ +CD₃ OD) δ: 1.80(s, 3H), 2.11(dd, 1H, J=5, 14 Hz), 2.26(s,3H), 2.64(s, 3H), 3.93(dd, 1H, J=7, 14 Hz), 4.03(s, 3H), 4.86(s, 2H),5.22(s, 2H), 6.99(dd, 1H, J=5, 7 Hz), 7.40-7.72(m, 4H), 7.80-8.08(m,2H), 9.04(s, 1H).

MS (m/e): 581 (M⁺).

REFERENCE EXAMPLE 7

In 30 ml of chloroform was dissolved 500 mg (0.86 mmol) of Compound i,and 0.64 ml (8.6 mmol) of ethanethiol and 199 mg (0.86 mmol) of camphorsulfonic acid were added to the solution, followed by stirring at roomtemperature for 2 hours. The reaction solution was washed successivelywith saturated sodium bicarbonate aqueous solution and saturated sodiumchloride aqueous solution and then dried over anhydrous magnesiumsulfate. The solvent was distilled off under reduced pressure, and theresidue was subjected to silica gel column chromatography (5% ethylacetate-toluene) to give 340 mg (63%) of Compound i as colorless prismsshowing a melting point of 181° to 184° C.

NMR (CDCl₃) δ: 1.28(t, 3H, J=8 Hz), 1.76(s, 3H), 2.11(dd, 1H, J=5, 14Hz), 2.26(s, 3H), 2.53(q, 2H, J=8 Hz), 2.80(s, 3H), 3.97(dd, J=7, 14Hz), 4.00(s, 2H), 4.01(s, 3H), 5.36(s, 2H), 7.02(dd, 1H, J=5, 7 Hz),7.14-7.80(m, 4H), 7.92-8.20(m, 2H), 9.13(s, 1H).

MS (m/e): 626 (M⁺).

REFERENCE EXAMPLE 8

In ethyl acetate was dissolved 125 mg (0.2 mmol) of Compound i, and 200mg of Raney nickel was added to the solution, followed by heating underreflux for 7 hours. The reaction solution was filtered through Celiteand the solvent was distilled off under reduced pressure to give 116 mg(100%) of Compound k as pale yellow powder.

NMR (CDCl₃) δ: 1.75(s, 3H), 2.04(dd, 1H, J-5, 14 Hz), 2.20(s, 3H),2.48(s, 3H), 2.61(s, 3H), 3.86(dd, 1H, J=7, 14 Hz), 3.99(s, 3H), 5.08(s,2H), 6.91(dd, 1H, J=5, 7 Hz), 7.16-7.64(m, 4H), 7.80-8.04(m, 2H),8.80(s, 1H).

MS (m/e): 566 (M⁺).

REFERENCE EXAMPLE 9

To a solution of 2.49 g (5.7 mmol) of Compound 20 in 30 ml of anhydrousTHF were added 2.70 g (14.2 mmol) of p-toluenesulfonyl chloride, 1.97 ml(14.2 mmol) of triethylamine and 0.69 g (5.7 mmol) ofN,N-dimethylaminopyridine, followed by stirring at room temperatureovernight. To the reaction mixture was added 100 ml of THF and theresulting solution was washed with an acid and an alkali. The solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (chloroform-methanol) to give 1.11 g(33%) of Compound l as pale yellow powder.

Melting point: 207°-210° C.

¹ H-NMR (DMSO-d₆ +CDCl₃) δ: 9.24(d, 1H, J=8 Hz), 8.15-7.8(m, 3H),7.65-7.2(m, 4H), 6.62(dd, 1H, J=5, 7 Hz), 4.95(d, 1H, J=10 Hz), 4.80(d,1H, J=10 Hz), 4.45(s, 2H), 3.05(dd, 1H, J=7, 14 Hz), 2.55(s, 3H),2.36(dd, 1H, J=5, 14 Hz), 2.12(s, 3H).

MS (m/z): 422 [M⁺ -167 (OTs)].

REFERENCE EXAMPLE 10

To 131 mg (0.3 mmol) of the free amine of Compound 30 were added 94 mg(0.45 mmol) of N-Cbz-glycine and 93 mg (0.45 mmol) ofdicyclohexylcarbodiimide (DCC), followed by stirring overnight. Aftercompletion of the reaction, the precipitates were separated by suctionfiltration. The solvent in the filtrate was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (2% MeOH-CHCl₃) to give 156 mg (83%) of Compound m aspale yellow powder showing a melting point of 157° to 162° C.

NMR (DMSO-d₆) δ: 1.96-2.30(m, 1H), 2.19(s, 3H), 2.58(s, 1H),2.72-3.08(m, 1H), 3.60-3.88(m, 4H), 5.00(s, 2H), 5.11(s, 2H), 5.72(s,1H), 6.86-8.20(m, 13H), 8.56(s, 1H), 9.20(dd, 1H, J=2, 8 Hz).

REFERENCE EXAMPLE 11

Compound n (61 mg, 32%) was obtained from 131 mg (0.3 mmol) of the freeamine of Compound 30 and 97 mg (0.45 mmol) of Boc-L-proline in a similarmanner as in Reference Example 10 as pale yellow powder showing amelting point of 203° to 213° C.

NMR (DMSO-d₆ +D₂ O) δ: 1.47(s, 9H), 1.66-2.32(m, 5H), 2.23)br, s. 3H),2.64-3.04(m, 1H), 3.20-3.64(m, 4H), 4.16-4.44(m, 1H), 4.98(br. s, 2H),6.88-7.60(m, 5H), 7.90-8.20(m, 3H), 9.16)br. d, 1H, J=8 Hz).

REFERENCE EXAMPLE 12

In 20 ml of THF and 10 ml of water was dissolved 438 mg (1 mmol) of thefree amine of Compound 30, and 420 mg (5 mmol) of sodiumhydrogencarbonate was added to the solution. Then, 0.21 ml (1.5 mmol) ofbenzyloxycarbonyl chloride was added thereto under ice cooling, followedby stirring at the same temperature for one hour. The reaction solutionwas washed with saturated NaCl aqueous solution and then dried overanhydrous magnesium sulfate. The solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (2% MeOH-CHCl₃) to give 282 mg (49.3%) of Compound o aspale yellow powder showing a melting point higher than 300° C.

NMR (DMSO-d₆) δ: 1.88-2.20(m, 1H), 2.15(s, 3H), 3.00(dd, 1H, J=7, 14Hz), 3.40-3.72(m, 2H), 5.00(br. s, 2H), 5.15(s, 2H), 5.60(s, 1H),5.68-6.96(m, 1H), 7.12-7.72(m, 10H), 7.90-8.16(m, 2H), 8.56(s, 1H),9.20(d, 1H, J=8 Hz).

REFERENCE EXAMPLE 13

Compound p (170 mg, 64.5%) was obtained from 262 mg (0.45 mmol) ofCompound o in a similar manner as in Example 65 as yellow powder showinga melting point of 281° to 285° C.

nNMR (CDCl₃ +DMSO-d₆) δ: 2.00-2.30(m, 1H), 2.18(s, 3H), 3.05(dd, 1H,J=7, 14 Hz), 3.48-3.90(m, 2H), 5.20(s, 2H), 5.69(s, 1H), 6.68-7.00(m,1H), 7.08-7.70(m, 10H), 8.01(d, 1H, J=7 Hz), 9.08(d, 1H, J=8 Hz),9.27(d, 1H, J=7 Hz).

We claim:
 1. A K-252 derivative represented by formula (I) and apharmacologically acceptable salt thereof ##STR30## wherein: R¹represents hydrogen, methyl, hydroxy, hydroxymethyl, lower alkoxy,bromine, chlorine or --NR⁵ R⁶ (wherein either R⁵ or R⁶ is hydrogen andthe other is hydrogen, carbamoyl, or lower alkylaminocarbonyl, or bothare lower alkyl) and R³ is hydrogen, or R¹ and R³ are the same andrepresent hydroxy, lower alkoxy or amino;R² is hydrogen or amino; R⁴ ishydrogen, chlorine, carbamoyl, lower alkyl, amino or --CH₂ CH₂ R⁷(wherein R⁷ is bromine, amino, di-lower alkylamino, hydroxy orhydroxy-substituted lower alkylamino); W₁ and W₂ are hydrogen or bothare combined together to represent oxygen; X is hydrogen, formyl, loweralkoxycarbonyl, ##STR31## (wherein R⁸ and R⁹ are independently hydrogen,lower alkyl or hydroxy-substituted lower alkyl, or R⁸ is hydrogen and R⁹is hydroxy), --CH₂ A {wherein A is hydroxy, azido, lower alkylthio,lower alkylsulfenyl, ##STR32## [wherein either R¹⁰ or R¹¹ is hydrogenand the other is hydrogen, lower alkyl, allyl, carboxylicacid-substituted lower alkyl, dihydroxy-substituted lower alkyl, aresidue of an α-amino acid in which the hydroxy of the carboxylic acidis removed or lower alkoxycarbonyl-substituted lower alkyl; or both arelower alkyl or chlorine-substituted lower alkyl; or R¹⁰ and R¹¹ arecombined together to form --CH₂ CH₂ --B--CH₂ CH₂ -- (wherein B is --CH₂--, --NH--, --S-- or --O--)], --N═CH--NMe₂ (wherein Me is methyl),--OCOCH₂ CH₂ CO₂ H or ##STR33## or --C═N--R¹² (wherein R¹² is hydroxy,amino, guanidino or 2-imidazolylamino); Y is hydroxy or carbamoyloxy; orX and Y are combined together to form, as --X--Y--, O═, --CH₂ O--, --CH₂OCOO--, --CH₂ --O--CS--0--, --CH_(2--NR) ¹³ --CO--O-- --CH₂--NH--CS--O--, --CH₂ --O--SO--O-- or ##STR34## (wherein R¹⁴ is loweralkyl or lower alkylthio); with the proviso that:when W₁ and W₂ arecombined together to represent oxygen, R¹, R² and R³ are all hydrogen;when R⁴ is lower alkyl, amino or --CH₂ CH₂ R¹, W₁ and W₂ are combinedtogether to represent oxygen; when Y is carbamoyloxy, R¹, R², R³, W₁ andW₂ are all hydrogen, R⁴ is carbamoyl and X is lower alkoxycarbonyl; whenR⁴ is chlorine, R¹, R², R³, W, and W₂ are all hydrogen and X is loweralkoxycarbonyl; when X is hydrogen, formyl, ##STR35## (wherein R^(8a)and R^(9a) are independently hydrogen, lower alkyl, hydroxy-substitutedlower alkyl) or CH₂ Aa (wherein the Aa representation is the same as theabove A representation excluding hydroxy and amino) of --CH═N--R¹², R¹,R², R³, R⁴, W₁ and W₂ are all hydrogen; when X is CONHOH, R¹, R², R³ andR⁴ are all hydrogen; when X is lower alkoxycarbonyl, R⁴ is hydrogen,chlorine, carbamoyl, lower alkyl or --CH₂ CH₂ R^(7a) (wherein R^(7a) isbromine or di-lower alkylamino); when A is aminomethyl, R¹, R² and R³are hydrogen, and when W₁ and W₂ are combined together to representoxygen, R⁴ is hydrogen or amino and when W₁ and W₂ are hydrogen, R⁴ ishydrogen; when X and Y are combined together to represent, as --X--Y--,--O--, --CH₂ --O--, --CH₂ --o--CO--O--, --CH₂ --O--CS--O--, --CH₂--NR^(13a) --CO--O-- (wherein the R^(13a) representation is the same asthe above R¹³ representation excluding hydrogen and lower alkyl), --CH₂--NH--CS--O--, --CH₂ --o--SO--o-- or ##STR36## R¹, R², R³, R⁴, W₁ and W₂are all hydrogen; when X and Y are combined together to represent, as--X--Y, --CH₂ --NR^(13b) --CO--O-- [wherein R^(13b) is hydrogen or loweralkyl), R¹, R², R³ and R⁴ are all hydrogen; when R¹ is methyl, hydroxy,hydroxymethyl, lower alkoxy, bromine, chlorine or --NR⁵ R⁶, R² and R⁴are hydrogen; when R² is amino, R¹ and R⁴ are hydrogen; and thecombinations wherein R¹, R², R³, R⁴, W₁ and W₂ are hydrogen, X ismethoxycarbonyl and Y is hydroxy are excluded.
 2. A compound representedby formula (Ia) and a pharmacologically acceptable salt thereof:##STR37## {wherein Xa is alkoxycarbonyl having 3 to 6 carbon atoms,##STR38## (wherein R⁸ and R⁹ are independently hydrogen, lower alkyl orhydroxy-substituted lower alkyl, or R⁸ is hydrogen and R⁹ is hydroxy),or --CH₂ Ab [wherein Ab is hydroxy, azido, lower alkylthio, loweralkylsulfenyl, ##STR39## (wherein either R^(10a) or R^(11a) is hydrogenand the other is hydrogen lower alkyl, or both are lower alkyl; orR^(10a) and R^(11a) are combined together to form --CH₂ CH_(2;l)--O--CH₂ CH₂ --)]; and Ya is hydroxy; or Xa and Ya are combined togetherto form, as --Xa--Ya--, --O--, --CH₂ --O--, --CH₂ O--CO--O--, --CH₂--o--CS--O--, --CH₂ --NH--CO--O-- or --CH₂ --o--SO--O--}.
 3. A K-252derivative represented by formula (Ib) and a pharmacologicallyacceptable salt thereof: ##STR40## [wherein R^(1a) represents hydrogen,methyl, hydroxymethyl, bromine, hydroxy, lower alkoxy or --NR⁵ R⁶(wherein either R⁵ or R⁶ is hydrogen and the other is hydrogen,carbamoyl or lower alkylaminocarbonyl, or both are lower alkyl) and R³is hydrogen, or R^(1a) and R³ are the same and represent hydroxy, loweralkoxy or amino; R^(4a) is hydrogen, chlorine, carbamoyl or lower alkyl;W₁ and W₂ are hydrogen or both are combined together to representoxygen; Xb is formyl, hydroxymethyl, lower alkoxycarbonyl, --CH═N--R¹²(wherein R¹² is hydroxy, amino, guanidino or 2-imidazolylamino), and Ybis hydroxy or carbamoyloxy, or Xb and Yb are combined together to form,as --Xb--Yb--, --CH₂ --NH--CS--O-- or ##STR41## (wherein R^(14a) islower alkylthio); with the priviso that: when W₁ and W₂ is oxygen, bothR^(1a) and R³ are hydrogen, Xb is lower alkoxycarbonyl or hydroxymethyl,R^(4a) is hydrogen or lower alkyl and Yb is hydroxy;when Yb iscarbamoyloxy, R^(1a), R³, W₁ and W₂ are all hydrogen, R^(4a) iscarbamoyl and Xb is lower alkoxycarbonyl; when R^(4a) is chlorine,R^(1a), R³, W₁ and W₂ are all hydrogen and Xb is lower alkoxycarbonyl;when R^(4a) is lower alkyl, R^(1a) and R³ are hydrogen, W₁ and W₂ areoxygen and Xb is lower alkoxycarbonyl or hydroxymethyl; when Xb isformul or --CH═N--R¹², R^(1a), R³, R^(4a), W₁ and W₂ are all hydrogen;when Xb is hydroxymethyl, R^(1a) and R³ are hydrogen and W₁ and W₂ areoxygen; when Xb and Yb are combined together to form, as --Xb--Yb--,##STR42## R^(1a), R³, R^(4a), W₁ and W₂ are all hydrogen; and thecombinations wherein R^(1a), R³, R^(4a), W₁ and W₂ are all hydrogen andXb is lower alkoxycarbonyl are excluded].
 4. A K-252 derivativerepresented by formula (Ic) and a pharmacologically acceptable saltthereof: ##STR43## wherein R^(1b) represents hydrogen, methyl, hydroxy,chlorine, hydroxymethyl, lower alkoxy, bromine or --NR⁵ R⁶ (whereineither R⁵ or R⁶ is hydrogen and the other is hydrogen, carbamoyl orlower alkylaminocarbonyl, or both are lower alkyl) and R^(3a) ishydrogen, or R^(1b) and R^(3a) are the same and represent hydroxy, loweralkoxy or amino;R² is hydrogen or amino; R^(4b) is hydrogen, amino or--CH₂ CH₂ R⁷ (wherein R⁷ is bromine, amino, di-lower alkylamino, hydroxyor hydroxy-substituted lower alkylamino); W₁ and W₂ are hydrogen or bothare combined together to represent oxygen; Xc is hydrogen, loweralkoxycarbonyl, hydroxyaminocarbonyl or CH₂ Ad {wherein Ad is hydroxy,##STR44## [wherein either R^(10b) or R^(11b) is hydrogen and the otheris hydrogen, allyl, carboxylic acid-substituted lower alkyl,dihydroxy-substituted lower alkyl, a residue of an α-amino acid in whichthe hydroxy of the carboxylic acid is removed or loweralkoxycarbonyl-substituted lower alkyl; or both are chlorine-substitutedlower alkyl; or R^(10b) and R^(11b) are combined together to form --CH₂CH₂ --Va--CH₂ CH₂ -- (wherein Ba is --CH₂ --, --NH-- or --S--)],--N═CH--NMe₂, ##STR45## Yc is hydroxy; or Xc and Yc are combinedtogether to form, as --Xc--Yc--, --CH₂ --NR^(13c) --CO;13 O-- [whereinR^(13c) is lower alkyl, allyl, formylmethyl, --CH₂ CH(OH--CH₂ OH,##STR46## (wherein R^(14b) is lower alkyl); with the proviso that:whenW₁ and W₂ are oxygen, R^(1b), R² and R^(3a) are all hydrogen; whenR^(4b) is amino or --CH₂ CH₂ R⁷, W₁ and W₂ are oxygen; when R^(1b) ismethyl, hydroxy, hydroxymethyl, lower alkoxy, bromine or --NR⁵ R⁶, R²and R^(4b) are hydrogen and Xc is hydroxymethyl; when R^(1b) ischlorine, R² and R^(4b) are hydrogen and Xc is lower alkoxycarbonyl orhydroxymethyl; when R² is amino, R^(1b) and R^(4b) are hydrogen and Xcis lower alkoxycarbonyl or hydroxymethyl; when Xc is hydrogen or CH₂ Ae(wherein the Ae representation is the same as the above Adrepresentation excluding hydroxy and amino), R^(1b), R², R^(3a), R^(3a),W₁ and W₂ are all hydrogen; when Xc is hydroxyaminocarbonyl, R^(1b), R²,R^(3a) and R^(4b) are all hydrogen and W₁ and W₂ are oxygen; when Xc islower alkoxycarbonyl and W₁ and W₂ are oxygen, R^(4b) is --CH₂ CH₂R^(7a) (wherein R^(7a) is bromine or di-lower alkylamino); when Xc isaminomethyl, R^(1b), R² and R^(3a) are hydrogen, W₁ and W₂ are oxygenand R^(4b) is hydrogen or amino; when Xc is hydroxymethyl and W₁ and W₂are oxygen, R^(4b) is amino or --CH₂ CH₂ R⁷ ; when Xc and Yx arecombined together to represent, as --Xc--Yc--, --CH₂ --NR^(13d)--CO--O-- (wherein the R^(13d) representation is the same as the aboveR^(13c) representation excluding lower alkyl) or ##STR47## R^(1b), R²,R^(3a), R^(4b), W₁ and W₂ are all hydrogen; when Xc and Yc are combinedtogether to represent, as --Xc--Yc--, --CH₂ --NR^(13e) --CO--O--(wherein R^(13e) is lower alkyl), R^(1b), R², R^(3a) and R^(4b) arehydrogen and W₁ and W₂ are oxygen; and the combinations wherein R^(1b),R², R^(3a), R^(4b), W₁ and W₂ are hydrogen, Xc is methoxycarbonyl and Ycis hydroxy and the combinations wherein R^(1b), R², R^(3a), R^(4b), W₁and W₂ are hydrogen, Xc is hydroxymethyl and Yc is hydroxy are removed.